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Project:Blood typing: Difference between revisions
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→PCR (Nicholas, Ben) 16/1/13
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== PCR (Nicholas, Ben) 16/1/13== | == PCR (Nicholas, Ben) 16/1/13== | ||
* Repeated PCR on successful samples from November 2012. | * Repeated PCR on successful samples from November 2012. | ||
* Ran PCR with PB1 primers on these two + 1 positive control with SYBR green | * Ran PCR with PB1 primers on these two + 1 positive control with SYBR green Taq readymix | ||
* 25ul total PCR reaction volume - 5ul template (1ul for PC), 2.5ul each forward & reverse primers, 12.5ul SYBR green | * 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. | ||
* PCR left overnight. 7ul of samples loaded, 7ul of ladder loaded. (Less than previous times) | * PCR left overnight. 7ul of samples loaded, 7ul of ladder loaded. (Less than previous times) | ||
