Berk2006-ConjugationTeam3

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JL_11/29/06

  1. ran gel:

(Lane 1=1, lane 2=2... etc. Lane 13= ladder)

JL_11/27/06

Tecan measurements

Culture Tecan measurement
TG1 42
J01122 4
J01022 2987
J23090 44
J23090-J23086 145
J23093 55
J23093-J23086 200

JL_11/20/06

  1. made -80 of [lock 3n]J23093
  2. plated [lock 3n]J23093
  3. grew up [lock 3n]J23093
  4. organized boxes in freezer

JL_10/13/06

Conjugated EC100d with

  1. J01064 db1K0 #1
  2. J01064 db1K0 #2
  3. J01064 db1K0 #3
  4. J01064 db1K0 #4
  5. J23015-J01064

on Tri Amp

Also I made TriK and TriA plates

JL_10/09/06

  1. Mate J23016 with Ec100D (see if the trbC knockout transfers its

plasmid, so plate on TriK)

  1. Do genomic minipreps of J23087 (4 clones)
  2. Mini and map J23086 (4 clones)
  3. Mini and map J23088 (2 clones)

JL_09/29/06

  1. Preperative digest of pJ23006-J23006 (AlwnI/Spei, 2068 Long)
  2. Preperative digest of psB1A2-J01003 (AlwnI/Xbai, 925 short)
  3. 4 Analytical digests(AvrII/SpeI) each of J23074 (want band at 23097) and J23075 (want band at 824)
  4. grow up J23016

JL_09/22/06

Today's to do list:

  1. stock and mini pSB1AG0-J23078, map EcoRI/SpeI
  2. grow up J23055 #1 for electro comp
  3. grow up J23016 (trbC::Cm) for the knockout (later gen. mini and PCR with JL16/17)
  4. Rlam/J23057 x pSB1A2-J01003 redo

JL_09/20/06

Today I... - mini-preped J23074 and J23075, and pSB1AKG0-b0015 - gen. mini and PCR J23055 - grew up Rlam/J23057 - analytical digest of J23074 and J23075


JCAnderson 22:35, 20 September 2006 (EDT)

Four clones were spotted the other day, all 4 were ampS and camS. The JL3/4 PCRs were done on genomic preps today. Jen setup and sam ran PCRs of J23055 clones 1 and 2. Clone 1 looks correct, it was stocked and replated to make comp cells.

JL_09/18/06

Today I have to:

  1. Grow up J23074 and J23075 (4 clones each)
  2. Grow and spot J23055 (43 degrees J23015 x pCP20)
  3. Run PCR of J23069 with your traG 20mers