Kun:LabNotes/ExonomeSeq/2007-12-20: Difference between revisions
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(New page: ==Exp.1 Cloning of the size selected fragments from 12/19/07== ===Blunting of the DNA fragment=== 5X Phusion buffer: 1ul 10mM dNTP: 0.25ul DNA: 3...) |
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==Exp.1 Cloning of the size selected fragments from 12/19/07== | ==Exp.1 Cloning of the size selected fragments from 12/19/07== | ||
PAGE size selected fragments: | |||
[[Image:2007-12-20-E55kv1-PAGE-selected-fragments.png|500px]] | |||
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=== | ===Blunting of the DNA fragment=== | ||
5X Phusion buffer: 1ul | 5X Phusion buffer: 1ul |
Revision as of 02:16, 21 December 2007
Exp.1 Cloning of the size selected fragments from 12/19/07
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
5X Phusion buffer: 1ul 10mM dNTP: 0.25ul DNA: 3.65ul Phusion: 0.1ul 72C 10min
Blunt-end ligation
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.