Team:IvyTech-South Bend/Notebook

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==Notebook==
==Notebook==
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{| cellpadding="20" align="center"
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=06 }}
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=07 }}
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=08 }}
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=09 }}
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=10 }}
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|{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=11 }}
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|}
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===IGEM Part #s and Uses===
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{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=06 }}{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=07 }}{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=08 }}
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BBa_T9002 – Producer Controler Device–Plate2 Well 9A
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BBa_I732017 – LacZ with our GFP 1—Plate 2 Well 3K
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BBa_F2620– LuxR with terminator– Plate 2 Well 6E
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BBa_I13522 –Tet Prop with GFP—Plate 2 Well 8A
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{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=09 }}{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=10 }}{{ #calendar: title=Talk:Team:IvyTech-South_Bend |year=2010 | month=11 }}
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BBa_T13521 Red Flouro tet RFP—Plate 2 Well 6O
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BBa_pSB1C3—Plasmid—Plate 1 Well 3A
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BBa_pSB1T3—Tetracycline resistant plasmid—Plate 1 Well 7A
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BBa_pSB1AT3--high copy number plasmid carrying ampicillin and tetracycline resistance--Plate 1 Well 13A
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 +
== 6/28/10 ==
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Preparing SOB
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== 6/30/10 ==
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Richard Lab:Electroporation of E. coli
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== 7/28/10 ==
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 +
Usage and extraction
== 8/24/10 ==
== 8/24/10 ==
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Today we will be pouring new plates for streaking new transformed cells.
Today we will be pouring new plates for streaking new transformed cells.
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We will be running a gel to determine if our DNA sample was successfully electroplated.
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We will be running a gel to determine if our DNA sample was successfully electroplated.
== 8/27/10 ==
== 8/27/10 ==
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Today we will be electroplating part KBBa_3131010
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Today we will be electroplating part KBBa_3131010
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== 8/31/10 ==
== 8/31/10 ==
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Results of transformation - the plates had growth but (no) distinct colony pattern.
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Results of transformation - the plates had growth but (no) distinct colony pattern.
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== 8/31/10 ==
== 8/31/10 ==
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After agar is dissolved, and allowed to cool, I will add Amp in different doses to specific plates. JH
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After agar is dissolved, and allowed to cool, I will add Amp in different doses to specific plates. JH
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== 9/1/10 ==
== 9/1/10 ==
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Today the team will be making up the same plates as I made yesterday. Professor T. wants new Amp stock made to document how much is used into the stock.
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Today the team will be making up the same plates as I made yesterday. Professor T. wants new Amp stock made to document how much is used into the stock.
== 9/2/10 ==
== 9/2/10 ==
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Results from streaking –
Results from streaking –
== 9/2/10 ==
== 9/2/10 ==
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Protocol For Making LB-Broth/Agar
Protocol For Making LB-Broth/Agar
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== 9/3/10 ==
== 9/3/10 ==
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Today I’m finishing making the LB/Agar from yesterday.
Today I’m finishing making the LB/Agar from yesterday.
== 9/9/10 ==
== 9/9/10 ==
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Due to conflict we will be changing from E Coli to yeast.
Due to conflict we will be changing from E Coli to yeast.
== 9/10/10 ==
== 9/10/10 ==
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Lux Casette Right Promoter BBa_I1051
Lux Casette Right Promoter BBa_I1051
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== 9/14/10 ==
== 9/14/10 ==
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Today we found growth from our Electroporated E – Coli parts
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Today we found growth from our Electroporated E – Coli parts
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== 9/15/10 ==
== 9/15/10 ==
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Today we will be extracting DNA from out electroporated cells/T9002
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- Today we will be extracting DNA from out electroporated cells/T9002
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B-galactosidase Assay
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- Wash our grown pos. bacteria with 10 percent glycerol to create Electro comp. cells
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- Then Electroporate T9002 into Elect. comp grown pos/t bacteria
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First we will be extracting DNA from BBa_T9002 Electroporated Cells using this protocol.
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== 9/15/10 ==
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I poured 10 mL LB – Lennox into 7 different 15 mL sterile Centrifuge tubes “2” for our electroporated parts T9002, K091107 with 10 mL Amp added 5 for our grown t/pos bacteria w/out Amp labeled then added one loop full of bacteria into each then placed on shaker overnight.
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Irene streaked our bacteria to set up for grown staining then placed into 37 degree incubation overnight.
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== 9/16/10 ==
== 9/16/10 ==
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Today we will wash
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Today we will wash our electrocomp grown t/pos bacteria with 1.2 mL Glycerol then electroporate
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Following protocol from page 15
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First prof. T sterilized the glycerol passing it through the .2 micron filter into 50 mL sterile centrifuge tube
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1 – then we added 1 mL 10 percent Glycerol to suspended cells
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2 – then centrifuged at 1000 rpm for 10 min
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- then removed supernate with BR 1000 micropipettor
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- then add 1 mL 10 percent Glycerol then repeat steps 1 & 2 “4” times then remove sup. and freeze cells at -80 degrees C freezer.
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- Today we also set up a new IGem work are in back lab look at pic
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== 9/17/10 ==
== 9/17/10 ==
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Today we will be doing a DNA Extraction from/of part T9002 for electroporation this will determine if we have a stable host for our beast this will also Work!
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Today we will be doing a DNA Extraction from/of part T9002 for electroporation this will determine if we have a stable host for our beast
 
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this will also Work!
 
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Following protocols from pgs 15 &19
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== 10/18/10 ==
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-instead of Electroporating our part we will use PG10 to determine if these lines are good hosts
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First we will be pouring a full pack of LB/Agar/Amp plates using the LB/Agar I made on 9/2/10 then adding amp made by DG on 9/1/10 we will also add 5 mL Arabonse to the Agar before pouring plates have been poured now letting stand for an hour to harden.
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cutting enzymes
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-Now I will be making 1L of LB/Broth for Culturing.
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== 9/21/10 ==
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Lot 9082989 ex. 2014-02-28
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BD Difco LB/Broth – Lennox 20 g per 1L
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so I will put 10 g into 2 elenmyier flask/capped one weighed 10.0023 g and the other weighed 10.0011 g then I added 500 mL into each flask then shook to mix then dissolved in microwaved for 8 oz setting once then placed into Autoclave for 15 min @ 121 degrees C until done.
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Now we are going electroporate our grown t/pos Electro Comp Cells from yesterday with PG10 using protocol from pg 15 changing # 5 to 5 mL of PG10
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Today we will electroporate Agrobacterium with part T9002
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We will be using a different protocol for electroporation of grown t/pos bacteria from Bio-Rad’s website prof. T will print and I’ll place here.
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The mycobacterium will be done by electroporation protocol on pg 15.
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Electrophoresis
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1) First I placed a Cuvette into Ice bath and thawed electro comp cells
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== 9/22/10 ==
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2) I set up 5 1.5 mL Sterile Centrifuge tubes  with 900 mL SOB media
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3) we will electroporate Irene’s electrocomp cells w/PG10 and mine will contain the parts after electroporation.
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Code
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Today we have growth from electroporation done yesterday we are going to test by hitting with (AHL) the 20 mL grew not 5 mL sample
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AT – A.tumefacions SL – S.lactis BM – B.magetanium MP – Mycobacteria
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BT – B.thurigensis
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split cells using 40 mL Hy part my cells then electroporate
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== 9/22/10 ==
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I electroporated all except mycobacteria now letting cells sit for at least
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The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens
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Just added 1 mL to electroporated Cultures they had sat for 20 min before adding.
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- Now I will electroporate mycobacteria using E – Coli protocol from pg 15 using 40 mL cells 4 mL pg10 Electroporation hit twice Read out 1.8 kv @ 5.2 ms then 1.79 kv @ 5.1 ms. then placed into 900 mL SOB media then incubate @ 30 degrees C for 3 hrs.
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Results
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== 9/23/10 ==
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From this we found that agrobacteria will be the best host for our beast/IGem
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we will do further testing towards our ultimate goal.
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 +
Today I have been setting up for the trial of our part T9002 inside agrobacteria to see if in the presence of AHL it floureces.
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== 9/21/10 ==
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==9/24/10 ==
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Today I’m testing absorbance of our trials @ 395nm
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Today we will electroporate  Agrobacterium with part T9002 using protocol from pg  22 – 23
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== 9/28/10 ==
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1) I will put frozen electro comp cells from -80 degrees C freezer
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Today we will be picking 2 colonies from the Lacz, LuxR electroporated plates that were streaked on 9/22/10
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2) We will do two “2” electroporations with (AT) one with 5 mL T9002 Gene, and one with 20 mL
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3) Icing Cuvette (.1) and now prepared to electroporate
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4) Reading for 20 mL was 2.2 kv @ 0.70 ms
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5) then removed with 1 mL SOB media
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6) Placed into 1.5 mL Sterile Centrifuge tube at room temp for 3 hrs prof T. will streak.
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 +
== 9/30/10==
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LacZ without GFP also with RBS and terminator
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== 9/22/10 ==
 
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==10/1/10==
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Today we have growth from electroporation done yesterday we are going to test by hitting with (AHL) the 20 mL grew
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electrophoresis
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not 5 mL sample
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1) Scrap bacteria from transformed plate
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2) Set up (3) Cuvettes  1 with 200 mL of bacteria + 800 mL LB – Lennox
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broth                            2 with 200 mL of bacteria + 800 mL of A.tumafaciens
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superant                        3 with 200 mL of bacteria + 800 mL of E – Coli Superant
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                                    4 with 200 mL LB broth + 800 mL of LB
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                                    5 with 200 mL LB broth + 800 mL of A tumafaciens
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      6 with 200 mL LB broth + 800 mL of E – Coli Superant
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change 500 mL cells either Agro-T9002 for sample A,B,C or Agro for D,E,F and 500 mL Sup + 200 mL SOB
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 +
==10/5/10==
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== 9/22/10 ==
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pTet GFP
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==10/7/10==
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The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens we will use suspended T9002 in agrobacteria prof T. set up yesterday as our 200 mL spl.
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Electroporating 2 parts with E-coli bacteria
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- Today I will be electroporating part #’s  I732094 (Lacz) – GFP, and part # F2620 (LuxR) into E – Coli Electrocomp cells.
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DG pulled the DNA parts from the Bio Brick and placed into 50 mL Electro comp E – Coli cells and 5 ml of DNA spls. now we will electroporate using protocol from pg 15  We recovered  cells shooting 900 mL into Cuvette then drawing it off with electroplated cells.
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Placed at room temp to grow for 1 hr then place and suspend in LB/Amp (10 mL)
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- I took our streaked to try to get an Isolated Colony Streaking and Heating between each pass. then placed into incubator.
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- Results after letting incubate for 2 days we have found that the incubator was set to too low of a temp we raised the temp to 37 degrees C from 26 degrees C we will see where we are at on Tuesday.
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 +
==10/8/10==
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== 9/23/10 ==
 
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LB-Lennox Broth & Electroporation
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Today I have been setting up for the trial of our part T9002 inside agrobacteria to see if in the presence of AHL it floureces.
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==10/12/10==
 +
hold until Friday
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1) I took and set up new growing Cultures of T9002 and E – Coli electro comp cells placed 5 mL into 50 mL sterile tubes one with Agro w/T9002 and one with E – Coli to make electro comp cells.
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==10/13/10==
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2) then drew off “2” 1 mL inciments of T9002 Sol. and placed 4.5 mL of Amp w/T9002
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Discoveries!
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3) now we are ready to perform trial taking Clean 1.5 mL Centrifuge tubes labeling 1 – 6 the first contains 200 mL  Argo/T9002 & 800 mL LB/Broth second w/200 mL Argo/T9002 w/800 mL Argobacteria Supernate. Third w/200 mL Argo/T9002 w/800 mL E – Coli Supernate (AHL) we are looking for a glow from this one Fourth 200 mL LB Broth – 800 mL LB Broth Fifth 200 mL LB Broth – 800 mL Argobacteria Sixth 200 mL LB Broth – 800 mL E – Coli Supernate
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Then they will all be tested inside a florimeter. Dylan is running a Spec reading on Agrobacteria/and Agro/w T9002 to see if the concentrations are close to the same. then do the trial he found that absorbance was 4 times higher so Dylan diluted the sample of T9002 ¼ adding LB – Lennox to this gave a spec reading of .303 and org. was .333 a difference of about 10 percent so we believe that will be ok we will work with this . See Dylans Lab notebook for more accurate details.
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for now I will be making and washing electrocomp E – Coli cells to be frozen
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==10/15/10==
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1) spinning off supernate from cells in 50 mL Centifuge
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2) then add 1 mL 10 percent glycerol and remove place in 1.5 mL centrifuge tube I will split cells into 6 centrifuge tubes adding 1 mL 10 percent glycerol and spin inside cold centrifuge @ 1000x for 10 min.
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3) then repeat a total of 4 times
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4) then leave about 40 mL of glycerol into the tubes and freeze @ -80 degrees C until needed.
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I took the electro comp cells I’m about to wash and ran a spec read I took 20 mL of E – Coli cells and placed into 980 mL of 10 percent glycerol then vortexed to mix then pipetted  on absorbance of .15 50
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Using openwetware.org protocol  
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20 mL into 1000w 1000/.20 = 50 X .15 = 7.5 so our OD is 7.5 mL
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so what I did was split the cells into 6 seperate tubes to be washed following protocol from pg20 while Dylan runs the Trial on T9002 from pg25 For results see pg 30
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==
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B-galactosidase Assay
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9/24/10 ==
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 +
BBL Trypticase soy Broth and Agar
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Today I’m testing absorbance of our trials @ 395nm
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DPBS
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1) .733 @ 395nm .678 @ 395nm
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2) .855 @ 395nm .586 @ 395nm
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3) .892 @ 395nm .693 @ 395nm
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4) .708 @ 395nm .733 @ 395nm
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5) .700 @ 395nm .700 @ 395nm
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6) .693 @ 395nm .729 @ 395nm
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Blank) .001 @ 395nm .001 @ 395nm
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1) I will blank the spec using LB/Lennox made by GM on 4/9/10
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== 10/19/10==
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2) –Change- I blanked using the LB – Lennox Broth w/200 mL of SOB media – both made by me.
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Protocol-- Electroporation man for E-Coli
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3) I took and used the same Cuvette for every spec scan washing with DI water between every trial
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== 10/21/10 ==
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4) Placing about 1 mL every trial
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High Efficiency Electrotransformation of E.coli
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5) Set spec to 395nm we believe that their was too much absorbance in trials 4,5, & 6 so we will do same procedure using florimider. Results say that the GFP is not being expressed in the presence of (AHL)/#3)
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- Prof T will now take sample ran in spec and run inside florimidor to check UV absorbance blanking with LB/SOB mixture.
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== 10/25/10 ==
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Now prof. T. is running the samples in the Fluo. he is blanking w/LB-SOB using 490ex and 510 EM filters Blank read out to .009 +/- .0006 10005 Fluo units for T9002 inside E – Coli
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Keeping Vol. the same for all spls. blank again before running spls. now running
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(A) (0) Fluo units we have no florescence
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(B) (160) Fluo units a little bit of florescence
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(C) (0) Fluo units this means GFP not producing
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(D) (0) Fluo units expected
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(E) (0) Fluo units expected
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(F) (0) Fluo units expected
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Results show no presence of GFP so we will break cells open then test again.
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DNA from BlueGuy
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== 9/28/10 ==
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Today we will be picking 2 colonies from the Lacz, LuxR electroporated plates that were streaked on 9/22/10 pg 26
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1) we will pick 2 colonies from each plate and transfer 15 mL LB – Lennox
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2) and 50 mL LB – Lennox with E – Coli & Agrobacteria growth. ( 2 tubes)
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3) poured (2) 40 mL tubes of LB then added 40 mL of Amp to them
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4) poured (4) 14 mL tubes of LB then added 14 mL of Amp to them
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Did all this inside Biological fume hood to keep sterile
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Now I will make LB – Lennox Broth (1L)
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1) Weigh out 10 g LB – Lennox Broth powder and place into 500 mL 18 ohm H2O and dissolve inside microwave using Beverage setting 8 oz then autoclave
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2) Repeat (2) twice for (1L)
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3) 1) 10.0266 g 2) 10.0698 g
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4) Placed each into 1L Capped Elenmyier flasks and added using a 500 mL grad cylinder filling with 500 mL 18 ohm H2O and adding 500 to each 1L flask
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5) Capped and microwaved on beverage setting then shook to mix
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-
6) Then placed in autoclave on liquid setting/ 121 degrees C for 15 min will place into fridge for use.
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 +
== 10/26/10 ==
 +
Final steps pack and ship
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''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
 
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Latest revision as of 00:44, 28 October 2010

Discussion

Contents

Notebook

June
MTWTFSS
  [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/1_June_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/2_June_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/3_June_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_June_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/5_June_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/6_June_2010&action=edit 6]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/7_June_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/8_June_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/9_June_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/10_June_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_June_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/12_June_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/13_June_2010&action=edit 13]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/14_June_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/15_June_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/16_June_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/17_June_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_June_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/19_June_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/20_June_2010&action=edit 20]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/21_June_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/22_June_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/23_June_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/24_June_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/25_June_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/26_June_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/27_June_2010&action=edit 27]
[http://2010.igem.org/Talk:Team:IvyTech-South_Bend/28_June_2010 28] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/29_June_2010 29] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/30_June_2010 30]
July
MTWTFSS
      [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/1_July_2010 1] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/2_July_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/3_July_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_July_2010&action=edit 4]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/5_July_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/6_July_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/7_July_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/8_July_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/9_July_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/10_July_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_July_2010&action=edit 11]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/12_July_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/13_July_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/14_July_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/15_July_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/16_July_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/17_July_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_July_2010&action=edit 18]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/19_July_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/20_July_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/21_July_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/22_July_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/23_July_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/24_July_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/25_July_2010&action=edit 25]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/26_July_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/27_July_2010&action=edit 27] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/28_July_2010 28] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/29_July_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/30_July_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/31_July_2010&action=edit 31]
August
MTWTFSS
            [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/1_August_2010&action=edit 1]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/2_August_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/3_August_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_August_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/5_August_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/6_August_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/7_August_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/8_August_2010&action=edit 8]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/9_August_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/10_August_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_August_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/12_August_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/13_August_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/14_August_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/15_August_2010&action=edit 15]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/16_August_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/17_August_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_August_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/19_August_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/20_August_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/21_August_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/22_August_2010&action=edit 22]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/23_August_2010&action=edit 23] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/24_August_2010 24] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/25_August_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/26_August_2010&action=edit 26] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/27_August_2010 27] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/28_August_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/29_August_2010&action=edit 29]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/30_August_2010&action=edit 30] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/31_August_2010 31]
September
MTWTFSS
    [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/1_September_2010 1] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/2_September_2010 2] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/3_September_2010 3] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_September_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/5_September_2010&action=edit 5]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/6_September_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/7_September_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/8_September_2010&action=edit 8] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/9_September_2010 9] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/10_September_2010 10] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_September_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/12_September_2010&action=edit 12]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/13_September_2010&action=edit 13] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/14_September_2010 14] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/15_September_2010 15] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/16_September_2010 16] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/17_September_2010 17] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_September_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/19_September_2010&action=edit 19]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/20_September_2010&action=edit 20] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/21_September_2010 21] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/22_September_2010 22] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/23_September_2010 23] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/24_September_2010 24] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/25_September_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/26_September_2010&action=edit 26]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/27_September_2010&action=edit 27] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/28_September_2010 28] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/29_September_2010&action=edit 29] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/30_September_2010 30]
October
MTWTFSS
        [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/1_October_2010 1] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/2_October_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/3_October_2010&action=edit 3]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_October_2010&action=edit 4] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/5_October_2010 5] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/6_October_2010 6] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/7_October_2010 7] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/8_October_2010 8] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/9_October_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/10_October_2010&action=edit 10]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_October_2010&action=edit 11] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/12_October_2010 12] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/13_October_2010 13] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/14_October_2010 14] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/15_October_2010 15] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/16_October_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/17_October_2010&action=edit 17]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_October_2010&action=edit 18] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/19_October_2010 19] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/20_October_2010 20] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/21_October_2010 21] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/22_October_2010 22] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/23_October_2010 23] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/24_October_2010 24]
[http://2010.igem.org/Talk:Team:IvyTech-South_Bend/25_October_2010 25] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/26_October_2010 26] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/27_October_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/28_October_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/29_October_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/30_October_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/31_October_2010&action=edit 31]
November
MTWTFSS
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/1_November_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/2_November_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/3_November_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/4_November_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/5_November_2010&action=edit 5] [http://2010.igem.org/Talk:Team:IvyTech-South_Bend/6_November_2010 6] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/7_November_2010&action=edit 7]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/8_November_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/9_November_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/10_November_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/11_November_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/12_November_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/13_November_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/14_November_2010&action=edit 14]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/15_November_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/16_November_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/17_November_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/18_November_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/19_November_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/20_November_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/21_November_2010&action=edit 21]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/22_November_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/23_November_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/24_November_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/25_November_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/26_November_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/27_November_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/28_November_2010&action=edit 28]
[http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/29_November_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=Talk:Team:IvyTech-South_Bend/30_November_2010&action=edit 30]

IGEM Part #s and Uses

BBa_T9002 – Producer Controler Device–Plate2 Well 9A

BBa_I732017 – LacZ with our GFP 1—Plate 2 Well 3K

BBa_F2620– LuxR with terminator– Plate 2 Well 6E

BBa_I13522 –Tet Prop with GFP—Plate 2 Well 8A

BBa_T13521 Red Flouro tet RFP—Plate 2 Well 6O

BBa_pSB1C3—Plasmid—Plate 1 Well 3A

BBa_pSB1T3—Tetracycline resistant plasmid—Plate 1 Well 7A

BBa_pSB1AT3--high copy number plasmid carrying ampicillin and tetracycline resistance--Plate 1 Well 13A

6/28/10

Preparing SOB

6/30/10

Richard Lab:Electroporation of E. coli

7/28/10

Usage and extraction

8/24/10

Today we will be pouring new plates for streaking new transformed cells.

8/27/10

We will be running a gel to determine if our DNA sample was successfully electroplated.

8/27/10

Today we will be electroplating part KBBa_3131010

8/31/10

Results of transformation - the plates had growth but (no) distinct colony pattern.

8/31/10

After agar is dissolved, and allowed to cool, I will add Amp in different doses to specific plates. JH

9/1/10

Today the team will be making up the same plates as I made yesterday. Professor T. wants new Amp stock made to document how much is used into the stock.

9/2/10

Results from streaking –

9/2/10

Protocol For Making LB-Broth/Agar

9/3/10

Today I’m finishing making the LB/Agar from yesterday.

9/9/10

Due to conflict we will be changing from E Coli to yeast.

9/10/10

Lux Casette Right Promoter BBa_I1051

9/14/10

Today we found growth from our Electroporated E – Coli parts

9/15/10

Today we will be extracting DNA from out electroporated cells/T9002 B-galactosidase Assay

9/16/10

Today we will wash

9/17/10

Today we will be doing a DNA Extraction from/of part T9002 for electroporation this will determine if we have a stable host for our beast this will also Work!


10/18/10

cutting enzymes

9/21/10

Today we will electroporate Agrobacterium with part T9002

Electrophoresis

9/22/10

Today we have growth from electroporation done yesterday we are going to test by hitting with (AHL) the 20 mL grew not 5 mL sample

9/22/10

The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens

9/23/10

Today I have been setting up for the trial of our part T9002 inside agrobacteria to see if in the presence of AHL it floureces.

9/24/10

Today I’m testing absorbance of our trials @ 395nm

9/28/10

Today we will be picking 2 colonies from the Lacz, LuxR electroporated plates that were streaked on 9/22/10

9/30/10

LacZ without GFP also with RBS and terminator


10/1/10

electrophoresis

10/5/10

pTet GFP

10/7/10

Electroporating 2 parts with E-coli bacteria

10/8/10

LB-Lennox Broth & Electroporation

10/12/10

hold until Friday

10/13/10

Discoveries!

10/15/10

Using openwetware.org protocol

B-galactosidase Assay

BBL Trypticase soy Broth and Agar

DPBS

10/19/10

Protocol-- Electroporation man for E-Coli

10/21/10

High Efficiency Electrotransformation of E.coli

10/25/10

DNA from BlueGuy

10/26/10

Final steps pack and ship


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