Phase 1: PCR Amplification of Genes
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'''A. Isolating genes for the quorum sensing mechanism from genomic ''B. subtilis'' DNA''' | '''A. Isolating genes for the quorum sensing mechanism from genomic ''B. subtilis'' DNA''' | ||
| - | + | 1. ComX - pheromone, 168bp | |
Primers: Fwd = CGGAATTCGCTCTAGATGCAAGACCTAATTAACTACTTTTTAAATTATCC | Primers: Fwd = CGGAATTCGCTCTAGATGCAAGACCTAATTAACTACTTTTTAAATTATCC | ||
Rev = GACTGCAGCTACTAGTTAATCACCCCATTGACGGGTTATTGG | Rev = GACTGCAGCTACTAGTTAATCACCCCATTGACGGGTTATTGG | ||
Notes: | Notes: | ||
Forward primer contains EcoRI and XbaI cut sites; Reverse primer contains SpeI and PstI cut sites | Forward primer contains EcoRI and XbaI cut sites; Reverse primer contains SpeI and PstI cut sites | ||
| - | Amplification of ComX alone was unsuccessful. However, ComQ and ComX are adjacent on the genome so we subsequently amplified them together | + | Amplification of ComX alone was unsuccessful. However, ComQ and ComX are adjacent on the genome so we subsequently amplified them together |
Revision as of 20:47, 23 October 2006
A. Isolating genes for the quorum sensing mechanism from genomic B. subtilis DNA
1. ComX - pheromone, 168bp
Primers: Fwd = CGGAATTCGCTCTAGATGCAAGACCTAATTAACTACTTTTTAAATTATCC
Rev = GACTGCAGCTACTAGTTAATCACCCCATTGACGGGTTATTGG
Notes:
Forward primer contains EcoRI and XbaI cut sites; Reverse primer contains SpeI and PstI cut sites
Amplification of ComX alone was unsuccessful. However, ComQ and ComX are adjacent on the genome so we subsequently amplified them together
