Kun:LabNotes/ASE/2007-11-6: Difference between revisions

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(New page: ===Exp. 1 PAGE purification of the eSNP84k probes=== ===Exp. 2 Continue on the second round of probe prep.=== Starting from the ethanol precipitated PCR amplicons from 800ul reactions. ...)
 
 
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===Exp. 1 PAGE purification of the eSNP84k probes===
===Exp. 1 PAGE purification of the eSNP84k probes===


      Lib #1:  10ng/ul(380nM) x 40ul
      Lib #2:  15ng/ul(580nM) x 40ul
      Lib #3:  5ng/ul(200nM) x 40ul
      Lib #4:  13ng/ul(500nM) x 40ul
      Lib #5:  12ng/ul(460nM) x 40ul


===Exp. 2 Continue on the second round of probe prep.===
===Exp. 2 Continue on the second round of probe prep.===
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The DNAs were resuspended in 100ul H2O. I added 10ul 10X NEBuffer 4, 4ul BciVI, 37C 3h, 75C 10min. I have one extra tube of Lib #5, which was included in the experiment.
The DNAs were resuspended in 100ul H2O. I added 10ul 10X NEBuffer 4, 4ul BciVI, 37C 3h, 75C 10min. I have one extra tube of Lib #5, which was included in the experiment.
Ethanol precipitation, resuspended in 45ul dH2O, added 5ul 10X lambda exo buffer, 3ul lambda exo, 37C 3h, 75C 10min, add 5ul USER, 37C overnight.

Latest revision as of 02:22, 8 November 2007

Exp. 1 PAGE purification of the eSNP84k probes[edit]

     Lib #1:  10ng/ul(380nM) x 40ul 
     Lib #2:  15ng/ul(580nM) x 40ul
     Lib #3:   5ng/ul(200nM) x 40ul
     Lib #4:  13ng/ul(500nM) x 40ul
     Lib #5:  12ng/ul(460nM) x 40ul

Exp. 2 Continue on the second round of probe prep.[edit]

Starting from the ethanol precipitated PCR amplicons from 800ul reactions.

The DNAs were resuspended in 100ul H2O. I added 10ul 10X NEBuffer 4, 4ul BciVI, 37C 3h, 75C 10min. I have one extra tube of Lib #5, which was included in the experiment.

Ethanol precipitation, resuspended in 45ul dH2O, added 5ul 10X lambda exo buffer, 3ul lambda exo, 37C 3h, 75C 10min, add 5ul USER, 37C overnight.