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From 2010.igem.org

Module I, DT and Fan's part:

Change the plasmid for part I. pUC19 is suitable for most of the restriction sites and it is highly copied inside the E. coli. Thus primers 1-N-up (or do) have changed as follow.

eGFP

 1-E-up	5’-CAGCGTCGACTAGGGATAACAGGGTAATAGGAGGTCACTGTAATGGTGAGCAAGGGCGAG-3’(SalI)
 1-E-do	5’-CCACAAGCTTTTACTTGTACAGCTCGTCCATGCCG-3’(HindIII)

mCherry

 1-M-up	5’-CAGCGTCGACTAGGGATAACAGGGTAATAGGAGGTCACTGTAATGGTGAGCAAGGGCGAG-3’(SalI)
 1-M-do	5’-CCAC GGATCC CTACTTGTACAGCTCGTCCATGCCG-3’(BamHI)

Kan

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

Chlr

 1-C-up	5'-CAGCGGTACCTAGGGATAACAGGGTAATCGGCATGGACGAGCTGTACAAGTAAGGAGGTTGCTAAA ATGGAGAAAAAAATCACTG-3'(KpnI)
 1-C-do	5'-CCACGAATTCATTACCCTGTTATCCCTAACGTTCGCTAATGCCGTTGACGGCCCTCATCGCAGTACTGTTGTATTCAT-3'(EcoRI)

Find that the SuperMix is powerful in running PCR. It’s great news.

Module I, YX and ZY's part:

We PCR amplified the Kanamycin resistance and Chloramphenicol resistance genes. We also amplified the replication origin from pTKS/CS. Later, we ligated the PCR products into T vector for storage.