Project:Sex typing with amelogenin/results-20120311 - Revision history

Czechton: /* 1a. Not extracting enough DNA */

2014-04-25T00:37:01Z

1a. Not extracting enough DNA

← Older revision		Revision as of 00:37, 25 April 2014
Line 25:		Line 25:
	#. Try to extract DNA from another source. This could get gruesome: likely sources are other bodily fluids (blood, stool, semen -- so, yeah, let's try to stick with buccal cells); or flesh.		#. Try to extract DNA from another source. This could get gruesome: likely sources are other bodily fluids (blood, stool, semen -- so, yeah, let's try to stick with buccal cells); or flesh.
	#. Be more rigorous about extracting DNA: use a hook rather than a pipette (though actually the pipette seemed fine)		#. Be more rigorous about extracting DNA: use a hook rather than a pipette (though actually the pipette seemed fine)
	#. Investigate more DNA-extraction methods ~~(see [[../extractionmethods]])~~		#. Investigate more DNA-extraction methods

	=== 1b. DNA is contaminated by proteins, or by other things which inhibit PCR. ===		=== 1b. DNA is contaminated by proteins, or by other things which inhibit PCR. ===

Czechton at 23:54, 24 April 2014

2014-04-24T23:54:51Z

@@ Line 1: / Line 1: @@
 Two separate DNA extraction protocols: Chelex and meat tenderiser.
@@ Line 10: / Line 11: @@
 I conclude that the DNA extraction technique is inadequate.
 [[Category:Biohacking]]

Montyphy: Montyphy moved page Sex typing with amelogenin/results-20120311 to Project:Sex typing with amelogenin/results-20120311

2013-06-03T15:28:36Z

Montyphy moved page Sex typing with amelogenin/results-20120311 to Project:Sex typing with amelogenin/results-20120311

← Older revision

Revision as of 15:28, 3 June 2013

Asoko at 20:42, 8 May 2013

2013-05-08T20:42:20Z

← Older revision		Revision as of 20:42, 8 May 2013
Line 10:		Line 10:

	I conclude that the DNA extraction technique is inadequate.		I conclude that the DNA extraction technique is inadequate.

			[[Category:Biohacking]]

Wzdd: Created page with "Two separate DNA extraction protocols: Chelex and meat tenderiser. Chelex: rinse mouth with salt. Spit small amount of liquid into tube. Add 500ul Chelex. Incubate at 98 degrees..."

2012-03-11T01:02:31Z

Created page with "Two separate DNA extraction protocols: Chelex and meat tenderiser. Chelex: rinse mouth with salt. Spit small amount of liquid into tube. Add 500ul Chelex. Incubate at 98 degrees..."

New page

Two separate DNA extraction protocols: Chelex and meat tenderiser.

Chelex: rinse mouth with salt. Spit small amount of liquid into tube. Add 500ul Chelex. Incubate at 98 degrees for 6 minutes. Centrifuge for 5 minutes. Take 10ul supernatant.

Meat tenderiser: rinse mouth with salt. Spit small amount of liquid into tube. Add equal amount of detergent. Vortex for 30 seconds. Add a few grains meat tenderiser. Vortex for 30 seconds. Allow to sit for several seconds. Add a small amount of rubbing alcohol. Observe DNA strands precipitating out of solution. Take 10ul of liquid and DNA strands. Incubate at 70 degrees for several minutes. Add 5 ul water.

PCR was set at D=94 degrees, 1 minute; A=57 degrees, 1 minute; E = 72 degrees, 1 minute. I used 5 ul of sample, 6 ul of primers, 12.5 ul of Taq master mix.

Result was visualised on 1% agarose gel stained with 2ul/40ml of ethidium bromide under 300nm UV. DNA ladder was visible. Both samples showed a faint blur running ahead of the ladder and no other information.

I conclude that the DNA extraction technique is inadequate.