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From 2010.igem.org

Module I, group 2(b)

PCR the fragments of the eGFP, mCherry, Kan and Chlr gene within the pBMN-PIB, pBS34, pKD13 and pKD3 plasmids. The primers using are shown as follow:

eGFP

 0-E-up
 5’-CAGCGTCGACTAGGGATAACAGGGTAATAGGAGGTCACTGTAATGGTGAGCAAGGGCGAG-3’(SalI)
 0-E-do
 5’-CCAC AAGCTT TTACTTGTACAGCTCGTCCATGCCG-3’(HindIII)

mCherry

 0-M-up	      
 5’-CAGCAAGCTTTAGGGATAACAGGGTAATAGGAGGTCACTGTAATGGTGAGCAAGGGCGAG-3’(HindIII)
 0-M-do        
 5’-CCAC GGATCC CTACTTGTACAGCTCGTCCATGCCG-3’(BamHI)

Kan

 0-K-up	
 5’-CAGCGGATCCTAGGGATAACAGGGTAATCGGCATGGACGAGCTGTACAAGTAAGGAGGTCACTGATGATTGAACAAGATGGATTGCAC-3’(BamHI)
 0-K-do	      
 5’-CCACGGTACCACGTTCGCTAATGCCGTTGACGGCCTCCCCTTATTAGAAGAACTCGTCAA-3’(KpnI)

Chlr

 0-C-up	
 5'-CAGCGGTACCTAGGGATAACAGGGTAATCGGCATGGACGAGCTGTACAAGTAAGGAGGTTGCTAAAATGGAGAAAAAAATCACTG-3'(KpnI)
 0-C-do	
 5'-CCACGAATTCATTACCCTGTTATCCCTAACGTTCGCTAATGCCGTTGACGGCCCTCATCGCAGTACTGTTGTATTCAT-3'(EcoRI)

Run a gel, cut it and gel extract.