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

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

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

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