Phase 1: PCR Amplification of Genes
From 2006.igem.org
A. Isolating genes for the quorum sensing mechanism from genomic B. subtilis DNA
- ComQX: ComQ = 900bp, required accesory protein for ComX production. ComX = 168bp, pheromone
- Primers:
- Fwd = CGGAATTCCCTCTAGATGAAGGAGATTGTGGAGCAAACATATTTAACG
- 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
- Series of DNA gel purifications (Xymo extraction) and PCR amplifications necessary to obtain sufficient quantity.
