Biomod/2011/Caltech/DeoxyriboNucleicAwesome/Protocols/GelPurification - Revision history

YAE LIM Lee at 07:13, 3 November 2011

2011-11-03T07:13:47Z

←Older revision		Revision as of 07:13, 3 November 2011
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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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YAE LIM Lee at 07:13, 3 November 2011

2011-11-03T07:13:21Z

←Older revision		Revision as of 07:13, 3 November 2011
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	== Gel Purification ==		== Gel Purification ==

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	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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YAE LIM Lee: New page: == 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 lan...

2011-11-03T03:41:59Z

New page: == 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 lan...

New page

== 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.