Kun:LabNotes/ExonomeSeq/2007-12-20
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Exp.1 Cloning of the size selected fragments from 12/19/07[edit]
PAGE size selected fragments: File:2007-12-20-E55kv1-PAGE-selected-fragments.png DNA concentration: GC1EP: 5.36ng/ul H2C: 1.64ng/ul 150bp: 3.73ng/ul 180/280bp 2.64ng/ul 350bp: 5.17ng/ul 470bp: too low to measure
Blunting of the DNA fragment[edit]
5X Phusion buffer: 1ul 10mM dNTP: 0.25ul DNA: 3.65ul Phusion: 0.1ul 72C 10min
Blunt-end ligation[edit]
pCR-Blunt 0.5ul Blunt PCR product 2ul 10X Ligation buffer 0.5ul Sterile water 1.5ul T4 DNA ligase (4U/ul) 0.5ul 16C 1h -> 65C 10min -> 4C hold.
Colony PCR[edit]
File:2007-12-23-Colony-PCR.png