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Project:Blood typing: Difference between revisions
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→Extraction and PCR (JJ, Bene, Will) 20-23/13
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==Extraction and PCR (JJ, Bene, Will) 20-23/13== | ==Extraction and PCR (JJ, Bene, Will) 20-23/13== | ||
* JJ and Bene extracted DNA from cheeks using [http://biology.clc.uc.edu/fankhauser/Labs/Genetics/Buccal_DNA_isolation/buccal_dna_images_index.html this] protocol. Small amount of cheek cells for each. | * JJ and Bene extracted DNA from cheeks on wed 20th using [http://biology.clc.uc.edu/fankhauser/Labs/Genetics/Buccal_DNA_isolation/buccal_dna_images_index.html this] protocol. Small amount of cheek cells for each. | ||
* Ran PCR with PB1 primers on this + a positive control. | * On 23rd Ran PCR with PB1 primers on this + a positive control. | ||
* 50ul total PCR reaction volume - 5ul dH20 (13ul for PC), 10ul template (2ul for PC), 5ul each forward & reverse primers, 25ul Taq readymix | * 50ul total PCR reaction volume - 5ul dH20 (13ul for PC), 10ul template (2ul for PC), 5ul each forward & reverse primers, 25ul Taq readymix | ||
* Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | * Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | ||
