Glasgow/Wetlab/Week10

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[[User:Christinemerrick|Christine]] realised that the site directed mutagenesis she had attempted on Thursday 30/08/07 was wrong.  S and Q were meant to be tested for (*a→d*), and D was meant to be tested for (*d→g*) but S and Q were tested for (*d→g*), and D was tested for (*a→d*).  This means the digests from 03/09/07 were also incorrect.  However, all is not lost because the sequencing results for S and D came back today and they show that both S and D contain (*d→g*).  This means that, theoretically, 1C, 1D and 1E from 03/09/07 should be positive, but this is not the case.
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[[User:Christinemerrick|Christine]] realised that the site directed mutagenesis she had attempted on Thursday 30/08/07 was wrong.  S and Q were meant to be tested for (*a→d*), and D was meant to be tested for (*d→g*) but S and Q were tested for (*d→g*), and D was tested for (*a→d*).  This means the digests from 03/09/07 were also incorrect.  However, all is not lost because the sequencing results for S and D came back today and they show that both S and D contain (*d→g*).  This means that, theoretically, digests of mutations 1C, 1D and 1E from 03/09/07 should be positive, but this is not the case.
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Before the sequencing results for S and D arrived, [[User:Christinemerrick|Christine]] set up site directed mutagenesis to correct that which she did 30/08/07.
=== Wednesday 5rd September 2007 ===
=== Wednesday 5rd September 2007 ===

Revision as of 14:37, 10 September 2007

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Contents

Week 10

Monday 3rd September 2007

  1. Christine's plates from 31/08/07 all grew except for 1B. Overnights were set up from 7 colonies on each of the plates on Sunday, labelled 1A1→1H7, 49 in total as there were none from plate 1B. Christine, Maia and Maija miniprpped them and digested them as follows:
    This table is incorrect, see 04/09/07.
    PCR tube Colony DNA is taken from Insert Enzyme Expected sizes Result
    1A D (*a→d*) EcoRI 3455bp, 18bp, 3654bp Negative
    1B D (*a→d*) PstI 274bp, 6553bp Negative
    1C S (*d→g*) EcoRI 7046bp, 18bp, 284bp Negative
    1D S (*d→g*) PstI 1155bp, 2511bp, 3682bp Negative
    1E S (*d→g*) PstI 1962bp, 1704bp, 3682bp Negative
    1F Q (*d→g*) EcoRI 7046bp, 18bp, 284bp Negative
    1G Q (*d→g*) PstI 1155bp, 2511bp, 3682bp Negative
    1H Q (*d→g*) PstI 1962bp, 1704bp, 3682bp Negative

    Tuesday 4nd September 2007

    1. Christine realised that the site directed mutagenesis she had attempted on Thursday 30/08/07 was wrong. S and Q were meant to be tested for (*a→d*), and D was meant to be tested for (*d→g*) but S and Q were tested for (*d→g*), and D was tested for (*a→d*). This means the digests from 03/09/07 were also incorrect. However, all is not lost because the sequencing results for S and D came back today and they show that both S and D contain (*d→g*). This means that, theoretically, digests of mutations 1C, 1D and 1E from 03/09/07 should be positive, but this is not the case.
    2. Before the sequencing results for S and D arrived, Christine set up site directed mutagenesis to correct that which she did 30/08/07.

      Wednesday 5rd September 2007

      Thursday 6th September 2007

      Friday 7th September 2007

      https://static.igem.org/mediawiki/2007/thumb/c/cc/Uog.jpg/50px-Uog.jpg Back To
      Glasgow's
      Main Page             		Back To
      Glasgow's
      Wetlab Log
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