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Project:Sex typing with amelogenin/results-20120212: Difference between revisions
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Project:Sex typing with amelogenin/results-20120212 (view source)
Revision as of 17:04, 12 February 2012
, 12 February 2012Experiment progress
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** Without master mix (?) | ** Without master mix (?) | ||
* PCR: Follow Wikipedia's basic melt, anneal, extend temperatures and times. | * PCR: Follow Wikipedia's basic melt, anneal, extend temperatures and times. | ||
Experiments to run: | |||
== Gel run 1 == | |||
1. PCR, no master mix, no template, just primers and 55 ul of buffer (Couldn't do this one, ran out of primers) | |||
2. PCR, no master mix, just template and primers and 5ul of buffer (Couldn't do this one, ran out of template and primers) | |||
3. PCR, master mix and primers, no template and 30 ul of buffer | |||
4. PCR, master mix, primers, and template (from top of tube) | |||
5. PCR, master mix, primers, and template (from bottom of tube) | |||
6. PCR, master mix (old one), primers, and template (from top of tube) | |||
7. PCR, master mix (old one), primers, and template (from bottom of tube) | |||
== DNA extraction == | |||
* 6ml of saline solution in mouth | |||
* swish for 10 seconds | |||
* spit back into tube | |||
* centrifuge for 5 minutes | |||
* pour off supernatant (liquid on top) into sink. | |||
* suspend tube in boiling water for 10 minutes | |||
* place on ice for 1 minute | |||
* centrifuge for 30 seconds | |||
* transfer 200ul of supernatant to fresh tube. This contains DNA. | |||
== Protocol == | |||
* 10 ul of primer (at a strength of 100 micromoles), x2 (so 10ul for each primer) | |||
* 25 ul of fast-start PCR Taq readymix | |||
* 30 ul of DNA. | |||
* 50 ul of mineral oil on top | |||
=== PCR === | |||
* D = 30 seconds at 98 degress | |||
* A = 40 seconds at 55 degrees (I think) | |||
* E = 60 seconds at 72 degrees | |||
* Repeat 25 times. | |||
* start: 14:22 | |||
* end: 16:02 | |||
=== Gel lanes (viewed with the gel oriented so the lanes are at the top) === | |||
1. Nothing | |||
2. PCR 3 above | |||
3. PCR 4 above | |||
4. PCR 5 above | |||
5. Ladder, 6ul | |||
6. PCR 6 above | |||
7. PCR 7 above | |||
8. Nothing | |||
Started electrophoresis at 16:11 at 100v. | |||
