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Project:Blood typing: Difference between revisions
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* No or not enough DNA extracted. Possibly fault of 'whirly' technique. Could also be due to leaving overnight. | * No or not enough DNA extracted. Possibly fault of 'whirly' technique. Could also be due to leaving overnight. | ||
* Poor resolution. Possibly due to new TBE. | * Poor resolution. Possibly due to new TBE. | ||
==Extraction and PCR (Will, Simon) 13/1/13== | |||
* Will and Simon extracted DNA from their cheeks using [http://biology.clc.uc.edu/fankhauser/Labs/Genetics/Buccal_DNA_isolation/buccal_dna_images_index.html this] protocol. (30 secs swishing and cheek chewing). Centrifuge for 5 minutes. Solution with chelex stirred every few minutes during 100C incubation. | |||
* Ran PCR with PB2 primers on this + a positive control. | |||
* 25ul total PCR reaction volume - 5ul template (1ul for PC), 2.5ul each forward & reverse primers, 12.5ul SYBR green Taq readymix, 2.5ul dH20 (6.5ul for PC) | |||
* Initial D - 96C 5 mins, then 35 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | |||
[[File:gel13jan2013.JPG|left|400px|thumb|none|Gel 13 Jan]] | |||
* PCR left overnight. 15ul of samples loaded, 6ul of ladder loaded. | |||
'''From left (after 35 mins at 80V:)''' | |||
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<br style="clear: both" /> | |||
'''Conclusions:''' | |||
