Thursday, July7

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PCR recipe Date: 7/7 Control A Control B Rx1 A Rx1 B Rx2 A Rx2 B Rx3 A Rx3 B Rx4 A Rx4 B Rx5 A Rx5 B Polymerase (pfu Ultra) 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul

10X buffer 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul 5ul

Forward primer (100ng/ul)1.25ul 1.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul

Reverse primer (100ng/ul)1.25ul 1.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul 6.25ul

Plasmid 2ul 2ul 0.5ul 0.5ul 1ul 1ul 5ul 5ul 10ul 10ul 15ul 15ul

dNTPs 1ul 1ul 1ul 1ul 1ul 1ul 1ul 1ul 1ul 1ul 1ul 1ul

Nuclease free water 34.5ul 34.5ul 26ul 26ul 25.5ul 25.5ul 21.5ul 21.5ul 16.5ul 16.5ul 11.5ul 11.5ul

this it the recipe for the pcr reaction of mutagenesis we did on july 7 thursday.

Cylcle                    temperature         time

Initial warmup 95 degrees 2 mins

Denaturation 95 degrees 30 sec

annealing 61.8 degrees 30 sec

extension 72 degrees 5min

final extension 72 degrees 10 min

store 4 degrees infinitive

Bivor