Difference between revisions of "Biomod/2011/Caltech/DeoxyriboNucleicAwesome/Protocols/GelPurification"

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- 240ul 10% APS
 
- 240ul 10% APS
 
- 24ul TEMED
 
- 24ul TEMED
    Note: APS should be used within 10 days
+
** Note: APS should be used within 10 days
  
 
4. Run the gel under 150V for roughly 4.5 hours ~ 6 hours (when the dye nearly runs off the gel) depending on the size of the sample. For example, if the length of DNA strand is 122nt~176nt, run for 6 hours. Change the buffer every 1.5 hours.
 
4. Run the gel under 150V for roughly 4.5 hours ~ 6 hours (when the dye nearly runs off the gel) depending on the size of the sample. For example, if the length of DNA strand is 122nt~176nt, run for 6 hours. Change the buffer every 1.5 hours.
  
 
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Latest revision as of 23:13, 2 November 2011

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Monday, December 18, 2017

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Gel Purification

1. Anneal the complex using a PCR machine

2. Wash the plates and dry for 45 minutes For 300ul sample, add 50ul BB_ND dye 170 μl of sample will be loaded per lane. Each sample will take 2 lanes, and one blank lane should be placed between different samples.

3. Make a 12% non-denaturing (ND) gel for purification Lulu Qian’s recipe for 40ml of 12% ND gel which works for one gel - 12ml 40% Acrylamide - 4ml 10x TAE/Mg++ - Fill with MQ water to 40ml total volume (add 24 ml) - 240ul 10% APS - 24ul TEMED

    • Note: APS should be used within 10 days

4. Run the gel under 150V for roughly 4.5 hours ~ 6 hours (when the dye nearly runs off the gel) depending on the size of the sample. For example, if the length of DNA strand is 122nt~176nt, run for 6 hours. Change the buffer every 1.5 hours.