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Project:Sex typing with amelogenin: Difference between revisions

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Project:Sex typing with amelogenin (view source)

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It's interesting in itself because we used a positive control thanks to Tom, plus known-good primers (also thanks to Tom). This suggests that the problem with intermittent results is the physical PCR process itself, rather than DNA extraction. (The previous good result already suggested that the primers and master mix weren't to blame.)
It's interesting in itself because we used a positive control thanks to Tom, plus known-good primers (also thanks to Tom). This suggests that the problem with intermittent results is the physical PCR process itself, rather than DNA extraction. (The previous good result already suggested that the primers and master mix weren't to blame.)


== More results ==
== 27th March 2012 ==


We've done several more tests since March 22, most of which seem to work. Summaries [http://wiki.london.hackspace.org.uk/view/Sex_typing_with_amelogenin/Results-latest here] The major problem seems to be quality of the extracted DNA: PCR product is frequently quite faint. As of April 26, 2012, DNA extraction technique will be the main focus for the amelogenin test (and for genetic testing of human DNA in general).
Meat tenderiser extraction: 35 cycles of D - 96° for 1:30, A - 55° for 2:00, E - 72° for 3:00. No result.
 
More detailed info in the lab book.
 
== 28th March 2012 ==
 
Expt. 1: Chelex extraction: 35 cycles of D - 96° for 1:00, A - 58° for 2:00, E - 72° for 3:00. No result.
 
Expt. 2: Chelex extraction: 35 cycles of D - 96° for 5:00, A - 45° for 5:00, E - 72° for 5:00. No result.
 
More detailed info in the lab book.
 
== 4th April 2012 ==
 
Mixed extractions: 35 cycles of D - 94° for 1:00, A - 50° for 0:30, E - 72° for 1:00. Mostly good results. Detailed info [https://lists.kentgeek.org/pipermail/lhs-biohacking/2012-April/000318.html here].
 
== 8th April 2012 ==
 
Meat tenderiser extraction: 35 cycles of D - 94° for 1:00, A - 55° for 1:00, E - 72° for 1:00. No result.
 
More detailed info in the lab book.
 
== 11th April 2012 ==
 
Mixed extractions: 35 cycles of D - 94° for 1:00, A - 54° for 0:30, E - 72° for 1:00. No result.
 
More detailed info in the lab book.
 
== 18th April 2012 ==
 
Chelex extraction: 35 cycles of D - 96° for 1:00, A - 56° for 0:30, E - 72° for 1:00. Good result. Only Nicolas' DNA used.
 
More detailed info in the lab book.
 
== 25th April 2012 ==
 
Chelex extraction: 35 cycles of D - 96° for 1:00, A - 56° for 0:30, E - 72° for 1:00. Good result. Detailed info [https://lists.kentgeek.org/pipermail/lhs-biohacking/2012-April/000352.html here].
 
== 16th May 2012 ==
 
Chelex extraction: 35 cycles of D - 96° for 1:00, A - 56° for 0:30, E - 72° for 1:00. No result.
 
More detailed info in the lab book.
 
== 19th June 2012 ==
 
Chelex extraction: 35 cycles of D - 96° for 1:00, A - 54° for 0:30, E - 72° for 1:00. No result.
 
More detailed info in the lab book.
 
== 1st August 2012 ==
 
Made a gel. 0.5g of agar. ~5g TBE. ~45g water (which sort?) 2.5ul.
 
More detailed info in the lab book.


= Complete protocol as of June 2012 =
= Complete protocol as of June 2012 =
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