Team:Cambridge/Lab book/Week 12

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Week: 3 4 5 6 7 8 9 10 11 12 13 14

Contents

Monday (10/09/12)

PCR of biobrick vector DNA


  • Attempted to run PCR of the backbone for the biobricks once more. Settings: Annealing temperature: 56°C, elongation time: 30secs.
  • After running on gel, saw that while the -8 vector amplification appears to be producing some correctly sized (~2kb) bands, the other two only seem to be forming many primer dimers. Analysis of the sequence of the prefix and suffix revealed a CG palindrome that appears to be causing self annealing. Inserts are not affected, as they have the palindrome at the 5' end of the primer.
  • Will retry at higher temperatures tomorrow.

Gibson assembly of biobricks


  • Attempted Gibson assembly for the -8 magnesium riboswitch biobrick with the backbone fragments produced earlier today and the insert fragments produced yesterday.

Transformation of e.coli with Gibson products


  • E.coli cells transformed with Gibson products produced earlier today.
  • Transformants plated out on 25μ/ml chloramphenicol plates.

Tuesday (11/09/12)

Wednesday (12/09/12)

Thursday (13/09/12)

Friday (14/09/12)

Saturday (15/09/12)

Sunday (16/09/12)