Berkeley LBL/Laina Notebook

Revision as of 05:16, 27 October 2007 by Silvapy (Talk | contribs)

Materials

Primer Design

Biobricking of bch-gene cluster and crtN gene.

Gene:

[[crtNgene]]

crtN

PCR of crtN on Heliobacillus mobilis G/C content on annealing region

crtN-F 40% BglII CrtN-R 50% BamHI and XhoI

Primers

crtN-F 3’ccaaaAGATCTgtgaaacatacagcaaaaaacctgggt5’

crtN-R 5’gctagCTCGAGttaGGATCCtcagtaactggctgacaagcct3’

bch B 3'ttaGGATCCccgttcgccttggtttgacttact5'

PCR

  PCR of bch-? genes using - 20ng/ul pHM6
  1ul pHM6 
  1ul dNTP
  5ul primer mix 
  0.5ul enzyme-phusion polymerase
  32.5ul H20
  50ul total

Dephosphorylation

Sequencing