Virginia Commonwealth/31 July 2009

From 2009.igem.org

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(New page: ===Results=== ''Maria and Afton'' * Overnight culture of I1352 grew, but no visible florescence * Transformation Results below {| cellpadding="10" align="center" border="0.9" padding="5" |...)
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==Tuesday 7 July 2009==
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===Results===
===Results===
''Maria and Afton''
''Maria and Afton''
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|-
! NEB 10 beta  
! NEB 10 beta  
-
| AMP || I1352 ligated to the backbone PSB4C5 || hundreds of colonies; some expressing RFP ||
+
| AMP || I1352 ligated to the backbone pSB4C5 || hundreds of colonies; some expressing RFP ||
|-
|-
! NEB 10 beta  
! NEB 10 beta  
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[[User:Trentay|Trentay]] 15:25, 31 July 2009 (UTC)
[[User:Trentay|Trentay]] 15:25, 31 July 2009 (UTC)
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* Interpretation of Transformation Results:
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** Shows the digestion/ligation process is not working
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** Because the I1352 grew successfully on an AMP plate after being 'ligated' with pSB4C5, shows that the original backbone is present
 +
*** So it was not properly digested
 +
** No growth was visible on the CM plate with the ligated I1352 with the CM resistant backbone pSB4C5
 +
*** If the digestion did not work properly, then the backbone could not be ligated with the I1352's promoter and RET
 +
** Conclusion:
 +
*** The enzymes are bad
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*** A new kit is being ordered
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*** Plans need to be made to repeat the Digestion and Ligation process with all promoters when kit arrives
 +
[[User:Trentay|Trentay]] 15:38, 31 July 2009 (UTC)
----
----
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 +
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===Tasks===
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''Afton and Maria''
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*
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----
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====Wetlab====
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''Afton and Maria''
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*

Revision as of 15:38, 31 July 2009

Contents

Tuesday 7 July 2009

Results

Maria and Afton

  • Overnight culture of I1352 grew, but no visible florescence
  • Transformation Results below
Name Plate Purpose Growth Observation
NEB 10 beta LB (+) Control shocked lawn
NEB 10 beta LB (+) Control lawn
NEB 10 beta AMP (-) Control no growth
NEB 10 beta CM (-) Control no growth
NEB 10 beta CM I1352 ligated to the backbone pSB4C5 no growth
NEB 10 beta AMP I1352 ligated to the backbone pSB4C5 hundreds of colonies; some expressing RFP
NEB 10 beta AMP I1352 ligated with itself hundreds of colonies; some expressing RFP

Trentay 15:25, 31 July 2009 (UTC)

  • Interpretation of Transformation Results:
    • Shows the digestion/ligation process is not working
    • Because the I1352 grew successfully on an AMP plate after being 'ligated' with pSB4C5, shows that the original backbone is present
      • So it was not properly digested
    • No growth was visible on the CM plate with the ligated I1352 with the CM resistant backbone pSB4C5
      • If the digestion did not work properly, then the backbone could not be ligated with the I1352's promoter and RET
    • Conclusion:
      • The enzymes are bad
      • A new kit is being ordered
      • Plans need to be made to repeat the Digestion and Ligation process with all promoters when kit arrives

Trentay 15:38, 31 July 2009 (UTC)



Tasks

Afton and Maria


Wetlab

Afton and Maria