IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-30

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(J36337 (A+B+C) transformants)
Current revision (00:21, 31 October 2006) (view source)
 
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We also inoculated 12 liquid cultures (2 mL LB Kan+Amp), one for each colony, from the restreak plates.
We also inoculated 12 liquid cultures (2 mL LB Kan+Amp), one for each colony, from the restreak plates.
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== Inoculation of J36834 (C), J36335 (A+C), J36336 (B+C) ==
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Inoculated 2 mL LB Kan+Amp of J36834 (C), J36335 (A+C), J36336 (B+C).

Current revision

Contents

Making stabs

  • Heat up LB agar 10min @ 50% power in microwave
  • Aliquot out 20mL
  • Add 40uL 50mg/uL AMP
  • Aliquot 1mL into each stab

Stabs made

 BBa_J   name           frozen stock
 36801   KaiA           2006_8_13KaiA
 36802   KaiB           2006_8_13KaiB
 36804   KaiC           2006_8_13KaiC
 36831   Lac+RBS+A      A9 green tube
 36832   Lac+RBS+B      colony E 10.21
 36834   Lac+RBS+C      M2-7 8.30
 36335   LacRBSALacRBSC colony 1 10.21
 36336   LacRBSBLacRBSC J36021 10/26/06

Updated biobricks registry & sent parts

yup

J36937 (A+B+C) transformants

We have 12 colonies, labeled A-L.

Restreaked A-L on two LB Kan plates.

Performed diagnostic PCR (8 uL PCR Supermix, 1 uL 2nM VF2, 1 uL 2nM VR primers) on each of the 12 colonies, along with a positive control (J36831 colony) and negative (no colony).

PCR schedule:

  • 95C for 15'00
  • Do 30 times:
    • 95C for 0'30
    • 55C for 0'30
    • 72C for 3'30
  • 72C for 10'00
  • 4C forever

PCR will finish around 0140.

We also inoculated 12 liquid cultures (2 mL LB Kan+Amp), one for each colony, from the restreak plates.

Inoculation of J36834 (C), J36335 (A+C), J36336 (B+C)

Inoculated 2 mL LB Kan+Amp of J36834 (C), J36335 (A+C), J36336 (B+C).

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