IGEM:MIT/2006/Notebook/2006-7-25

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IAA Smell Test

Added .1, .5, 1, 5, 10ul to 5ml liquid cultures of Top10 .5ul was detectable, but .1ul (which we might get) was less detectable

  • Tested today in preferred IK strains

Indole Knockouts

Out of 3 strains, the JC12337 did not smell as nice

  • note, previous disappointment with IK stemmed from contaminated LB

Made glycerols of MB408 and MEB61

osmY-Trunc Eco/Spe Digest

  • 5 uL Buffer 2
  • 0.5 uL BSA
  • 2 uL ~800 ng/uL Annealed osmY-Trunc primers
  • 1.25 uL EcoRI
  • 1.25 uL SpeI

ATF1A and R0030 Digest

Order ATF1-A mutation primers

Primer pair 2

                                   *
   Forward: 5' CACCCCCTGGATAAGCGAATTTGACATGAATGATAACAAAG 3'
   Reverse: 5' CTTTGTTATCATTCATGTCAAATTCGCTTATCCAGGGGGTG 3'
                                   *
    GC content: 41.46%           Location: 214-254
    Melting temp: 79.8°C         Mismatched bases: 1
    Length: 41 bp                Mutation: Substitution
    5' flanking region: 20 bp    Forward primer MW: 12619.37 Da
    3' flanking region: 20 bp    Reverse primer MW: 12587.31 Da