So I am wrangling with getting these literature based PKS I primers working for my PhD project. Finally success!!!
PKS I Primers
degKS2R 5’- GTICCIGTICCRTGISCYTCIAC
760 bp fragment
Schirmer, A., R. Gadkari, C. D. Reeves, F. Ibrahim, E. F. DeLong, and C. R. Hutchinson. 2005. Metagenomic analysis reveals diverse polyketide synthase gene clusters in microorganisms associated with the marine sponge Discodermia dissoluta. Appl Environ Microbiol 71:4840-4849.
Master mix for one reaction (bring up to 15 uL total with DNA and water)
PCR Buffer 1.5
Primers 0.15 each
DMSO 1.4 ***I typically don’t use DMSO so swap in water here)
GEL!!!! The first well in each line is the positive control