Virginia Commonwealth/4 August 2009

From 2009.igem.org

(Difference between revisions)
(Tuesday 4 August 2009)
(Tuesday 4 August 2009)
Line 27: Line 27:
** T4 Ligase Buffer (10 µL)
** T4 Ligase Buffer (10 µL)
** T4 Ligase (5 µL)
** T4 Ligase (5 µL)
 +
[[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC)
''Maria and Afton''
''Maria and Afton''
Line 33: Line 34:
* pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
* pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
* Transformation was done using DB 3.1
* Transformation was done using DB 3.1
 +
{| cellpadding="10" align="center" border="0.9" padding="5"
 +
|+
 +
|-
 +
! Name !! Plate !! Growth Observation !!
 +
|-
 +
! pSB1C3
 +
| CM||  ||
 +
|-
 +
! (+) Control
 +
| LB ||  ||
 +
|-
 +
! (+) Control zapped 
 +
| LB ||  ||
 +
|-
 +
! (-) Control
 +
| CM ||  ||
 +
|}
 +
[[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC)

Revision as of 22:33, 4 August 2009

Contents

Tuesday 4 August 2009

Results

Maria and Afton

  • Overnight culture of I1352 plated on AMP:
    • Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink
    • Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP
  • There is a possibility the pSB4C5 backbone may be a bad part
    • previous years it has not functioned properly
    • Gel electrophoresis indicates DNA Gel Purification may be necessary
  • Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing
    • They will be aliquoted into smaller amounts

Trentay 22:25, 4 August 2009 (UTC)


Tasks

Maria and Afton

  • Run a Gel Electrophoresis
  • Transform and grow up pSB1C3 with the death gene P1010

Trentay 22:11, 4 August 2009 (UTC)


Wetlab

  • New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts
    • XbaI (10 µL)
    • SpeI (5 µL)
    • PstI (10 µL)
    • EcoRI-HF (10 µL)
    • BSA (5 µL)
    • T4 Ligase Buffer (10 µL)
    • T4 Ligase (5 µL)

Trentay 22:33, 4 August 2009 (UTC)

Maria and Afton

  • Gel Electrophoresis was run
    • pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA
  • pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
  • Transformation was done using DB 3.1
Name Plate Growth Observation
pSB1C3 CM
(+) Control LB
(+) Control zapped LB
(-) Control CM

Trentay 22:33, 4 August 2009 (UTC)