Virginia Commonwealth/23 July 2009

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''Craig and Clay''
''Craig and Clay''
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* Electrotransformation of the limonene parts was reattempted.  Fifty microliters were streaked on each ampicillin-selective plate.
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* Electrotransformation of the limonene parts was reattempted.  Fifty microliters were streaked on each ampicillin-selective plate.  Note that SOC was used instead of LB after the electrotransformation.

Latest revision as of 15:09, 24 July 2009

May
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[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_May_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_May_2009&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/6_May_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/7_May_2009&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/8_May_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/9_May_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/10_May_2009&action=edit 10]
[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_May_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_May_2009&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/13_May_2009&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/14_May_2009&action=edit 14] [http://2009.igem.org/Virginia_Commonwealth/15_May_2009 15] [http://2009.igem.org/Virginia_Commonwealth/16_May_2009 16] [http://2009.igem.org/Virginia_Commonwealth/17_May_2009 17]
[http://2009.igem.org/Virginia_Commonwealth/18_May_2009 18] [http://2009.igem.org/Virginia_Commonwealth/19_May_2009 19] [http://2009.igem.org/Virginia_Commonwealth/20_May_2009 20] [http://2009.igem.org/Virginia_Commonwealth/21_May_2009 21] [http://2009.igem.org/Virginia_Commonwealth/22_May_2009 22] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/23_May_2009&action=edit 23] [http://2009.igem.org/Virginia_Commonwealth/24_May_2009 24]
[http://2009.igem.org/Virginia_Commonwealth/25_May_2009 25] [http://2009.igem.org/Virginia_Commonwealth/26_May_2009 26] [http://2009.igem.org/Virginia_Commonwealth/27_May_2009 27] [http://2009.igem.org/Virginia_Commonwealth/28_May_2009 28] [http://2009.igem.org/Virginia_Commonwealth/29_May_2009 29] [http://2009.igem.org/Virginia_Commonwealth/30_May_2009 30] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/31_May_2009&action=edit 31]
June
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[http://2009.igem.org/Virginia_Commonwealth/1_June_2009 1] [http://2009.igem.org/Virginia_Commonwealth/2_June_2009 2] [http://2009.igem.org/Virginia_Commonwealth/3_June_2009 3] [http://2009.igem.org/Virginia_Commonwealth/4_June_2009 4] [http://2009.igem.org/Virginia_Commonwealth/5_June_2009 5] [http://2009.igem.org/Virginia_Commonwealth/6_June_2009 6] [http://2009.igem.org/Virginia_Commonwealth/7_June_2009 7]
[http://2009.igem.org/Virginia_Commonwealth/8_June_2009 8] [http://2009.igem.org/Virginia_Commonwealth/9_June_2009 9] [http://2009.igem.org/Virginia_Commonwealth/10_June_2009 10] [http://2009.igem.org/Virginia_Commonwealth/11_June_2009 11] [http://2009.igem.org/Virginia_Commonwealth/12_June_2009 12] [http://2009.igem.org/Virginia_Commonwealth/13_June_2009 13] [http://2009.igem.org/Virginia_Commonwealth/14_June_2009 14]
[http://2009.igem.org/Virginia_Commonwealth/15_June_2009 15] [http://2009.igem.org/Virginia_Commonwealth/16_June_2009 16] [http://2009.igem.org/Virginia_Commonwealth/17_June_2009 17] [http://2009.igem.org/Virginia_Commonwealth/18_June_2009 18] [http://2009.igem.org/Virginia_Commonwealth/19_June_2009 19] [http://2009.igem.org/Virginia_Commonwealth/20_June_2009 20] [http://2009.igem.org/Virginia_Commonwealth/21_June_2009 21]
[http://2009.igem.org/Virginia_Commonwealth/22_June_2009 22] [http://2009.igem.org/Virginia_Commonwealth/23_June_2009 23] [http://2009.igem.org/Virginia_Commonwealth/24_June_2009 24] [http://2009.igem.org/Virginia_Commonwealth/25_June_2009 25] [http://2009.igem.org/Virginia_Commonwealth/26_June_2009 26] [http://2009.igem.org/Virginia_Commonwealth/27_June_2009 27] [http://2009.igem.org/Virginia_Commonwealth/28_June_2009 28]
[http://2009.igem.org/Virginia_Commonwealth/29_June_2009 29] [http://2009.igem.org/Virginia_Commonwealth/30_June_2009 30]
July
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    [http://2009.igem.org/Virginia_Commonwealth/1_July_2009 1] [http://2009.igem.org/Virginia_Commonwealth/2_July_2009 2] [http://2009.igem.org/Virginia_Commonwealth/3_July_2009 3] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_July_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_July_2009&action=edit 5]
[http://2009.igem.org/Virginia_Commonwealth/6_July_2009 6] [http://2009.igem.org/Virginia_Commonwealth/7_July_2009 7] [http://2009.igem.org/Virginia_Commonwealth/8_July_2009 8] [http://2009.igem.org/Virginia_Commonwealth/9_July_2009 9] [http://2009.igem.org/Virginia_Commonwealth/10_July_2009 10] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_July_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_July_2009&action=edit 12]
[http://2009.igem.org/Virginia_Commonwealth/13_July_2009 13] [http://2009.igem.org/Virginia_Commonwealth/14_July_2009 14] [http://2009.igem.org/Virginia_Commonwealth/15_July_2009 15] [http://2009.igem.org/Virginia_Commonwealth/16_July_2009 16] [http://2009.igem.org/Virginia_Commonwealth/17_July_2009 17] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/18_July_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/19_July_2009&action=edit 19]
[http://2009.igem.org/Virginia_Commonwealth/20_July_2009 20] [http://2009.igem.org/Virginia_Commonwealth/21_July_2009 21] [http://2009.igem.org/Virginia_Commonwealth/22_July_2009 22] [http://2009.igem.org/Virginia_Commonwealth/23_July_2009 23] [http://2009.igem.org/Virginia_Commonwealth/24_July_2009 24] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/25_July_2009&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/26_July_2009&action=edit 26]
[http://2009.igem.org/Virginia_Commonwealth/27_July_2009 27] [http://2009.igem.org/Virginia_Commonwealth/28_July_2009 28] [http://2009.igem.org/Virginia_Commonwealth/29_July_2009 29] [http://2009.igem.org/Virginia_Commonwealth/30_July_2009 30] [http://2009.igem.org/Virginia_Commonwealth/31_July_2009 31]
August
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[http://2009.igem.org/Virginia_Commonwealth/3_August_2009 3] [http://2009.igem.org/Virginia_Commonwealth/4_August_2009 4] [http://2009.igem.org/Virginia_Commonwealth/5_August_2009 5] [http://2009.igem.org/Virginia_Commonwealth/6_August_2009 6] [http://2009.igem.org/Virginia_Commonwealth/7_August_2009 7] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/8_August_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/9_August_2009&action=edit 9]
[http://2009.igem.org/Virginia_Commonwealth/10_August_2009 10] [http://2009.igem.org/Virginia_Commonwealth/11_August_2009 11] [http://2009.igem.org/Virginia_Commonwealth/12_August_2009 12] [http://2009.igem.org/Virginia_Commonwealth/13_August_2009 13] [http://2009.igem.org/Virginia_Commonwealth/14_August_2009 14] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/15_August_2009&action=edit 15] [http://2009.igem.org/Virginia_Commonwealth/16_August_2009 16]
[http://2009.igem.org/Virginia_Commonwealth/17_August_2009 17] [http://2009.igem.org/Virginia_Commonwealth/18_August_2009 18] [http://2009.igem.org/Virginia_Commonwealth/19_August_2009 19] [http://2009.igem.org/Virginia_Commonwealth/20_August_2009 20] [http://2009.igem.org/Virginia_Commonwealth/21_August_2009 21] [http://2009.igem.org/Virginia_Commonwealth/22_August_2009 22] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/23_August_2009&action=edit 23]
[http://2009.igem.org/Virginia_Commonwealth/24_August_2009 24] [http://2009.igem.org/Virginia_Commonwealth/25_August_2009 25] [http://2009.igem.org/Virginia_Commonwealth/26_August_2009 26] [http://2009.igem.org/Virginia_Commonwealth/27_August_2009 27] [http://2009.igem.org/Virginia_Commonwealth/28_August_2009 28] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/29_August_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/30_August_2009&action=edit 30]
[http://2009.igem.org/Virginia_Commonwealth/31_August_2009 31]
September
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  [http://2009.igem.org/Virginia_Commonwealth/1_September_2009 1] [http://2009.igem.org/Virginia_Commonwealth/2_September_2009 2] [http://2009.igem.org/Virginia_Commonwealth/3_September_2009 3] [http://2009.igem.org/Virginia_Commonwealth/4_September_2009 4] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_September_2009&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/6_September_2009&action=edit 6]
[http://2009.igem.org/Virginia_Commonwealth/7_September_2009 7] [http://2009.igem.org/Virginia_Commonwealth/8_September_2009 8] [http://2009.igem.org/Virginia_Commonwealth/9_September_2009 9] [http://2009.igem.org/Virginia_Commonwealth/10_September_2009 10] [http://2009.igem.org/Virginia_Commonwealth/11_September_2009 11] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_September_2009&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/13_September_2009&action=edit 13]
[http://2009.igem.org/Virginia_Commonwealth/14_September_2009 14] [http://2009.igem.org/Virginia_Commonwealth/15_September_2009 15] [http://2009.igem.org/Virginia_Commonwealth/16_September_2009 16] [http://2009.igem.org/Virginia_Commonwealth/17_September_2009 17] [http://2009.igem.org/Virginia_Commonwealth/18_September_2009 18] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/19_September_2009&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/20_September_2009&action=edit 20]
[http://2009.igem.org/Virginia_Commonwealth/21_September_2009 21] [http://2009.igem.org/Virginia_Commonwealth/22_September_2009 22] [http://2009.igem.org/Virginia_Commonwealth/23_September_2009 23] [http://2009.igem.org/Virginia_Commonwealth/24_September_2009 24] [http://2009.igem.org/Virginia_Commonwealth/25_September_2009 25] [http://2009.igem.org/Virginia_Commonwealth/26_September_2009 26] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/27_September_2009&action=edit 27]
[http://2009.igem.org/Virginia_Commonwealth/28_September_2009 28] [http://2009.igem.org/Virginia_Commonwealth/29_September_2009 29] [http://2009.igem.org/Virginia_Commonwealth/30_September_2009 30]
October
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      [http://2009.igem.org/Virginia_Commonwealth/1_October_2009 1] [http://2009.igem.org/Virginia_Commonwealth/2_October_2009 2] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/3_October_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_October_2009&action=edit 4]
[http://2009.igem.org/Virginia_Commonwealth/5_October_2009 5] [http://2009.igem.org/Virginia_Commonwealth/6_October_2009 6] [http://2009.igem.org/Virginia_Commonwealth/7_October_2009 7] [http://2009.igem.org/Virginia_Commonwealth/8_October_2009 8] [http://2009.igem.org/Virginia_Commonwealth/9_October_2009 9] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/10_October_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_October_2009&action=edit 11]
[http://2009.igem.org/Virginia_Commonwealth/12_October_2009 12] [http://2009.igem.org/Virginia_Commonwealth/13_October_2009 13] [http://2009.igem.org/Virginia_Commonwealth/14_October_2009 14] [http://2009.igem.org/Virginia_Commonwealth/15_October_2009 15] [http://2009.igem.org/Virginia_Commonwealth/16_October_2009 16] [http://2009.igem.org/Virginia_Commonwealth/17_October_2009 17] [http://2009.igem.org/Virginia_Commonwealth/18_October_2009 18]
[http://2009.igem.org/Virginia_Commonwealth/19_October_2009 19] [http://2009.igem.org/Virginia_Commonwealth/20_October_2009 20] [http://2009.igem.org/Virginia_Commonwealth/21_October_2009 21] [http://2009.igem.org/Virginia_Commonwealth/22_October_2009 22] [http://2009.igem.org/Virginia_Commonwealth/23_October_2009 23] [http://2009.igem.org/Virginia_Commonwealth/24_October_2009 24] [http://2009.igem.org/Virginia_Commonwealth/25_October_2009 25]
[http://2009.igem.org/Virginia_Commonwealth/26_October_2009 26] [http://2009.igem.org/Virginia_Commonwealth/27_October_2009 27] [http://2009.igem.org/Virginia_Commonwealth/28_October_2009 28] [http://2009.igem.org/Virginia_Commonwealth/29_October_2009 29] [http://2009.igem.org/Virginia_Commonwealth/30_October_2009 30] [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/31_October_2009&action=edit 31]

Contents

Thursday 23 July 2009

Results

Maria and Afton

  • All overnight cultures grew except one vial of NEB 10 beta seed stock
    • there was a second vial of seed stock that did grow successfully
  • Transformations were successful
Name Type Plate Purpose Growth Observation
NEB 10 beta seed stock LB make more seed stock lawn
NEB 10 beta electro comp. LB (+) Control/ make more stock lawn
NEB 10 beta electro comp. LB (+) Control shocked lawn
DB 3.1 electro comp. LB (+) Control shocked lawn
DB 3.1 electro comp. LB (+) Control/make more stock lawn
NEB 10 beta electro comp. Cm (-) Control no growth
NEB 10 beta electro comp. Cm J23100 w/ J06702 6 small colonies

Trentay 17:19, 23 July 2009 (UTC)

Tasks

Maria and Afton

  • Make cryogenic stocks of all overnight cultures
  • Make stocks of parts I1352 (RFP) and I13522 (GFP)
  • Update notebook, documents
  • Pick colonies from transformation
    • pick controls as well to make more seed and electrocompotent stock

Trentay 17:25, 23 July 2009 (UTC) Kevin and Adam

  • We need to PCR DNA padding onto the first nine synthesized promoters. When we ordered these oligo's we did not consider that we needed additional overhang from the BioBrick prefix and suffix in order for the restriction enzymes to work properly. Doing this PCR will correct that error. Bussingkm 20:56, 23 July 2009 (UTC)

Craig and Clay

  • Electrotransformation of the limonene pathway parts will be attempted a third time.

Wetlab

Maria and Afton

  • Cryogenic stocks were made
  • Overnight culture was made of J23100 w/ J06702 RET in LB+Cm

Trentay 21:25, 23 July 2009 (UTC)

Kevin and Adam

  • First we reconstitute the DNA. The rule of thumb is that you take 1/2 the amount of oligo's (nmol) of H2O in uL for ~100x DNA stock solution. Store in (-20C)
    • A problem was encountered while reconstituting the DNA. For several of the sequences the amount of water described above was not sufficient to reconstitute the DNA. It was too viscus and could not be pipetted at all. We doubled the amount of water that was used and made note of the approximate concentration.
## ~nmol DNA uL H2O Concentration
Design 1 25 nmol 12.5 uL 100x
Design 2 25 nmol 25 uL 50x
Design 3 25 nmol 25 uL 50x
Design 4 25 nmol 12.5 uL 100x
Design 5 25 nmol 25 uL 50x
Design 6 25 nmol 25 uL 50x
Design 7 25 nmol 25 uL 50x
Design 8 25 nmol 25 uL 50x
Design 9 25 nmol 25 uL 50x
Forward end padding 48.3 nmol 24.15 uL 100x
Reverse end padding 36.4 nmol 18.2 uL 100x
  • This DNA was frozen in Adam & Kevin's stock box in the vials that the DNA was shipped in.
  • Each sample of DNA was diluted to 1x and 100 uL with the appropriate amount of nanopure H2O to be used for PCR.
  • The first round of PCR was done using the standard protocol. There was an error when the machine was started. It somehow was paused while the temperature was 95C. This was discovered long after it occurred. Deng advised us to start over. We both have prior commitments and thus cannot stay late enough to start over. We will start over early in the morning. dNTP is made and mixed in a 1:1:1:1 ratio.

-Bussingkm 21:04, 23 July 2009 (UTC)

Craig and Clay

  • Electrotransformation of the limonene parts was reattempted. Fifty microliters were streaked on each ampicillin-selective plate. Note that SOC was used instead of LB after the electrotransformation.
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