Matt:LabNotes/2013-6-26

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Revision as of 18:15, 26 June 2013 by >Mzcai (Created page with "===Page Size Selection of 0 gap probes=== USER/DpnII digested 0 gap probes: Noi made 11.36ug/400ul [http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-5-29] ...")
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Page Size Selection of 0 gap probes

USER/DpnII digested 0 gap probes:

 Noi made 11.36ug/400ul [1]
 I made 10.58ug/180ul [2]

To run ~3ug/gel, I use 100ul of probes that Noi made for each of the 4 gels

  • Ran TBU gel at 220V for 30 min with samples and ladder prepared below
    • Heated TBE buffer in microwave
    • Denatured samples/ladder for 10 min at 75C before putting on cold rack for 1 min
    • Size selected for brightest band below 200bp
Components 1X Volume 4X Volume
Enzyme digested probes (~12.6ug/400ul) 105 420
TBE-Urea Buffer 2X 105 420
H2O 0 0
Total 210 840
Components 1X Volume 4X Volume
10-bp DNA ladder (1 ug/ul) 1 4
TBE-Urea Buffer 2X 5 20
H2O 4 16
total 10 40