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''' | ||
#ComX - pheromone, 168bp | #ComX - pheromone, 168bp | ||
| - | ::Primers: Fwd = CGGAATTCGCTCTAGATGCAAGACCTAATTAACTACTTTTTAAATTATCC | + | ::*Primers: Fwd = CGGAATTCGCTCTAGATGCAAGACCTAATTAACTACTTTTTAAATTATCC |
Rev = GACTGCAGCTACTAGTTAATCACCCCATTGACGGGTTATTGG | Rev = GACTGCAGCTACTAGTTAATCACCCCATTGACGGGTTATTGG | ||
| + | ::*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 20:53, 23 October 2006
A. Isolating genes for the quorum sensing mechanism from genomic B. subtilis DNA
- 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
