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*Testing of J37016, S01656
*Finish ligation for J37022 & electroporate (has it been finished yesterday?)
*Finish ligation for Dr. Frays aiiA into the T hanging plasmid & electroporate
*Culture J37036 & J37018 from plate in incubator for maxi & mini on Wednesday
*Culture T9002 from plate in incubator for new testing & making frozen stocks

Western Blot

  • There was no discernable difference in the proteins produced by the induced and non-induced cells
  • This indicates that there is something wrong with the mechanism involving S01656 expression, possibly its promoter
  • This does not however mean that the aiiA is wrong necessarily - testing of J37022 should clarify this.


Testing of S01656

  • Was started with the revised protocol but after results from Western Blot came out, was abandonded
  • Is not going to be tested any more since no protein seems to be transcribed at all

AiiA Ligation/electroporation

  • Samples run on gel and adequate bands cut out.
  • Glass milk purified and ligated into T9 overhanging vector
  • Forms New J37023
  • Electroporated in the afternoon


  • The small PCR fragment is not being amplified

Testing S01656 & J37016

  • Testing was not conducted as western blot indicates that there may be something wrong with the S01656 expression mechanism.
  • J37016 was being cultured for the S01656 testing. This test, too, was not finished since we are no longer carrying out testing of S01656. We have 3 good sets of data for J37016 so far.


Part Primer used Melting temp (Tm)
J37015RS Riboswitch 44[[:Category:{{{1}}}|{{{1}}}]]
J37020 Plasmid A2 reverse primer 56[[:Category:{{{1}}}|{{{1}}}]]
J37023 T3 and T7 primers (from Dr.Mann) 39[[:Category:{{{1}}}|{{{1}}}]] for both
J37025 Plasmid A2 forward primer 50[[:Category:{{{1}}}|{{{1}}}]]


  • Cultured up 4 colonies of J37036 in 110ml of LB Amp for maxi and mini
  • Cultured up 2 colonies of J37018 in 110ml of LB Amp for maxi and mini
  • Cultured up 2 colonies of T9002 in 2ml of LB Amp for tesing?
  • Cultured up J37015 in 2ml of LB Amp for testing
  • Cultured up J37016 in 2ml of LB Amp for testing