Team:IvyTech-South Bend/Notebook

From 2010.igem.org

(Difference between revisions)
(→9/21/10)
(→9/21/10)
 
(34 intermediate revisions not shown)
Line 48: Line 48:
==Notebook==
==Notebook==
 +
<html>
 +
<style type="text/css">
 +
table.calendar          { margin: 0; padding: 10px; }
 +
table.calendar td      { margin: 0; padding: 2px; vertical-align: top; }
 +
table.month .heading td { padding:2px; background-color:#0f6666; color:#00ff00; text-align:center; font-size:120%; font-weight:bold; }
 +
table.month .dow td    { color:#00ff00; text-align:center; font-size:110%; }
 +
table.month td.today    { background-color:#ddd;}
 +
table.month td {
 +
    border: none;
 +
    margin: 0;
 +
    padding: 1.5pt 2pt;
 +
    font-weight: bold;
 +
    font-size: 10pt;
 +
    text-align: center;
 +
    background-color: #000000;
 +
    }
 +
#bodyContent table.month a { background:none; padding:0 }
 +
.day-active { color:#00FF00 }
 +
.day-empty  { color:#006666 }
 +
</style>
 +
</html>
 +
{| cellpadding="20" align="center"
 +
|{{ #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 }}
 +
|{{ #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 }}
 +
|}
 +
===IGEM Part #s and Uses===
-
{{ #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 }}
+
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
-
{{ #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 }}
+
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 ==
== 8/24/10 ==
 +
Today we will be pouring new plates for streaking new transformed cells.
Today we will be pouring new plates for streaking new transformed cells.
Line 62: Line 113:
-
We will be running a gel to determine if our DNA sample was successfully electroplated.
+
We will be running a gel to determine if our DNA sample was successfully electroplated.
== 8/27/10 ==
== 8/27/10 ==
-
 
+
Today we will be electroplating part KBBa_3131010
-
Today we will be electroplating part KBBa_3131010
+
== 8/31/10 ==
== 8/31/10 ==
-
 
+
Results of transformation - the plates had growth but (no) distinct colony pattern.
-
Results of transformation - the plates had growth but (no) distinct colony pattern.
+
== 8/31/10 ==
== 8/31/10 ==
-
 
+
After agar is dissolved, and allowed to cool, I will add Amp in different doses to specific plates. JH
-
After agar is dissolved, and allowed to cool, I will add Amp in different doses to specific plates. JH
+
== 9/1/10 ==
== 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.
+
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 ==
-
 
Results from streaking –
Results from streaking –
== 9/2/10 ==
== 9/2/10 ==
-
 
Protocol For Making LB-Broth/Agar
Protocol For Making LB-Broth/Agar
-
 
-
 
== 9/3/10 ==
== 9/3/10 ==
-
 
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 ==
-
 
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 ==
-
 
Lux Casette Right Promoter BBa_I1051
Lux Casette Right Promoter BBa_I1051
Line 112: Line 153:
== 9/14/10 ==
== 9/14/10 ==
-
 
+
Today we found growth from our Electroporated E – Coli parts
-
Today we found growth from our Electroporated E – Coli parts
+
== 9/15/10 ==
== 9/15/10 ==
 +
Today we will be extracting DNA from out electroporated cells/T9002
 +
B-galactosidase Assay
-
- Today we will be extracting DNA from out electroporated cells/T9002
+
== 9/16/10 ==
 +
Today we will wash
 +
== 9/17/10 ==
-
== 9/16/10 ==
+
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!
-
Today we will wash our electrocomp grown t/pos bacteria with 1.2 mL Glycerol then electroporate
+
== 10/18/10 ==
-
Following protocol from page 15
+
-
First prof. T sterilized the glycerol passing it through the .2 micron filter into 50 mL sterile centrifuge tube
+
-
1 – then we added 1 mL 10 percent Glycerol to suspended cells
+
-
2 – then centrifuged at 1000 rpm for 10 min
+
-
- then removed supernate with BR 1000 micropipettor
+
-
- 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.
+
-
- Today we also set up a new IGem work are in back lab look at pic
+
cutting enzymes
 +
== 9/21/10 ==
-
== 9/17/10 ==
+
Today we will electroporate  Agrobacterium with part T9002
 +
Electrophoresis
-
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
+
== 9/22/10 ==
-
this will also Work!
+
-
Following protocols from pgs 15 &19
+
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
-
-instead of Electroporating our part we will use PG10 to determine if these lines are good hosts
+
-
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.
+
== 9/22/10 ==
-
-Now I will be making 1L of LB/Broth for Culturing.
+
The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens
-
Lot 9082989 ex. 2014-02-28
+
-
BD Difco LB/Broth – Lennox 20 g per 1L
+
-
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.
+
-
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
+
== 9/23/10 ==
-
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.
+
-
The mycobacterium will be done by electroporation protocol on pg 15.
+
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.
-
1) First I placed a Cuvette into Ice bath and thawed electro comp cells
+
==9/24/10 ==
-
2) I set up 5 1.5 mL Sterile Centrifuge tubes  with 900 mL SOB media
+
-
3) we will electroporate Irene’s electrocomp cells w/PG10 and mine will contain the parts after electroporation.
+
-
Code
+
Today I’m testing absorbance of our trials @ 395nm
-
AT – A.tumefacions SL – S.lactis BM – B.magetanium MP – Mycobacteria
+
-
BT – B.thurigensis
+
-
split cells using 40 mL Hy part my cells then electroporate
+
== 9/28/10 ==
-
I electroporated all except mycobacteria now letting cells sit for at least
+
Today we will be picking 2 colonies from the Lacz, LuxR electroporated plates that were streaked on 9/22/10
-
Just added 1 mL to electroporated Cultures they had sat for 20 min before adding.
+
-
+
-
- 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.
+
-
Results
+
== 9/30/10==
-
From this we found that agrobacteria will be the best host for our beast/IGem
+
LacZ without GFP also with RBS and terminator
-
we will do further testing towards our ultimate goal.
+
-
== 9/21/10 ==
+
==10/1/10==
 +
electrophoresis
-
Today we will electroporate  Agrobacterium with part T9002 using protocol from pg  22 – 23
+
==10/5/10==
-
== 9/22/10 ==
+
pTet GFP
 +
==10/7/10==
-
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
+
Electroporating 2 parts with E-coli bacteria
-
== 9/22/10 ==
+
==10/8/10==
-
The only one that should glow should be (C)/(3) E – Coli w/transformed part in LB For Supernate of A.Tumafaciens
+
LB-Lennox Broth & Electroporation
-
== 9/23/10 ==
+
==10/12/10==
 +
hold until Friday
 +
==10/13/10==
 +
Discoveries!
-
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.
+
==10/15/10==
-
==9/24/10 ==
+
Using openwetware.org protocol
 +
B-galactosidase Assay
-
Today I’m testing absorbance of our trials @ 395nm
+
BBL Trypticase soy Broth and Agar
-
== 9/28/10 ==
+
DPBS
 +
== 10/19/10==
 +
Protocol-- Electroporation man for E-Coli
 +
== 10/21/10 ==
 +
High Efficiency Electrotransformation of E.coli
-
Today we will be picking 2 colonies from the Lacz, LuxR electroporated plates that were streaked on 9/22/10  
+
== 10/25/10 ==
 +
DNA from BlueGuy
 +
== 10/26/10 ==
 +
Final steps pack and ship
Line 213: Line 252:
|-
|-
-
|[[Image:IvyTech-South_Bend_logo.png|200px|right|frame]]
+
|align="center"|[[Image:IvyTech-South_Bend_logo.png|200px|right|frame]]
|-
|-
|
|
-
''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
 
|
|
-
|align="center"|[[Team:IvyTech-South_Bend | Team Example]]
+
|
|}
|}

Latest revision as of 00:44, 28 October 2010

Discussion

Contents

Notebook

June
MTWTFSS
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
July
MTWTFSS
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
August
MTWTFSS
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31
September
MTWTFSS
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30
October
MTWTFSS
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
November
MTWTFSS
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 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


Your team picture
IvyTech-South Bend logo.png


Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety
Retrieved from "http://2010.igem.org/Team:IvyTech-South_Bend/Notebook"