Matt:LabNotes/2013-6-26: Difference between revisions

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>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] ...")
 
>Mzcai
mNo edit summary
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| align="center" style="background:#f0f0f0;"|'''4X Volume'''
| align="center" style="background:#f0f0f0;"|'''4X Volume'''
|-
|-
| Enzyme digested probes (~12.6ug/400ul)||105||420
| Enzyme digested probes (~11.36/400ul)||100||400
|-
|-
| TBE-Urea Buffer 2X||105||420
| TBE-Urea Buffer 2X||100||400
|-
|-
| H2O||0||0
| H2O||0||0
|-
|-
| Total||210||840
| Total||200||800
|-
|-
|  
|  

Revision as of 18:16, 26 June 2013

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 (~11.36/400ul) 100 400
TBE-Urea Buffer 2X 100 400
H2O 0 0
Total 200 800
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