Team:IvyTech-South Bend/Notebook

From 2010.igem.org

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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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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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- 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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== 9/23/10 ==
== 9/23/10 ==

Revision as of 13:42, 12 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]

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


9/16/10

Today we will wash our electrocomp grown t/pos bacteria with 1.2 mL Glycerol then electroporate 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


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!

Following protocols from pgs 15 &19 -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.

-Now I will be making 1L of LB/Broth for Culturing. 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 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.

1) First I placed a Cuvette into Ice bath and thawed electro comp cells 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 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

I electroporated all except mycobacteria now letting cells sit for at least 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 From this we found that agrobacteria will be the best host for our beast/IGem we will do further testing towards our ultimate goal.


9/21/10

Today we will electroporate Agrobacterium with part T9002 using protocol from pg 22 – 23

1) I will put frozen electro comp cells from -80 degrees C freezer 2) We will do two “2” electroporations with (AT) one with 5 mL T9002 Gene, and one with 20 mL 3) Icing Cuvette (.1) and now prepared to electroporate 4) Reading for 20 mL was 2.2 kv @ 0.70 ms 5) then removed with 1 mL SOB media 6) Placed into 1.5 mL Sterile Centrifuge tube at room temp for 3 hrs prof T. will streak.


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 1) Scrap bacteria from transformed plate 2) Set up (3) Cuvettes 1 with 200 mL of bacteria + 800 mL LB – Lennox broth 2 with 200 mL of bacteria + 800 mL of A.tumafaciens superant 3 with 200 mL of bacteria + 800 mL of E – Coli Superant 

                                    4 with 200 mL LB broth + 800 mL of LB
                                    5 with 200 mL LB broth + 800 mL of A tumafaciens

6 with 200 mL LB broth + 800 mL of E – Coli Superant 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


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



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