. Author manuscript; available in PMC: 2010 Jan 1.

Published in final edited form as: Nat Protoc. 2009;4(2):206–223. doi: 10.1038/nprot.2008.227

Table 3.

PCR primers and possible source of template for drug cassette amplification^*

Gene Cassette	Source	Primer Sequence	Annealing Temperature (°C)	Size (kb)
ampicillin (amp)	pBluescript SK(+) (Stratagene)	5' CATTCAAATATGTATCCGCTC 3' 5' AGAGTTGGTAGCTCTTGATC 3'	53	1.2
chloramphenicol (cat)	pPCR-Script Cam (Stratagene)	5' TGTGACGGAAGATCACTTCG 3' 5' ACCAGCAATAGACATAAGCG 3'	53	0.86
kanamycin (kan)	Tn5	5' TATGGACAGCAAGCGAACCG 3' 5' TCAGAAGAACTCGTCAAGAAG 3'	55	0.95
tetracycline (tetA)	Tn10	5' TCCTAATTTTTGTTGACACTCTA 3' 5' CTCTTGGGTTATCAAGAGGG 3'	55	1.34

When amplified with these primer pairs, all drug cassettes will contain a promoter and all but kan will have a transcriptional terminator.