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Project:Blood typing: Difference between revisions
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* 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. | * 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 | * Ran PCR with PB2 primers on these two + 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) | * 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. | * Initial D - 96C 5 mins, then 35 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | ||
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* PCR left overnight. 15ul of samples loaded, 6ul of ladder loaded. | * PCR left overnight. 15ul of samples loaded, 6ul of ladder loaded. | ||
'''From left (after 35 mins at 80V:)''' | '''From left (after 35 mins at 80V:)''' | ||
*1: | *1: 100-3000bp ladder. Ladder [http://www.nbsbio.co.uk/product.asp?pID=8951&cID=33 key]. | ||
*2: | *2: Simon PCR product | ||
*3: | *3: Will PCR product | ||
*4: | *4: PC PCR product | ||
*5: | *5: Simon gDNA | ||
*6: | *6: Will gDNA | ||
<br style="clear: both" /> | <br style="clear: both" /> | ||
'''Conclusions:''' | '''Conclusions:''' | ||
