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Project:Blood typing: Difference between revisions
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adding 16 Jan PCR
Mycoplasma (talk | contribs) |
Simulacrum (talk | contribs) (adding 16 Jan PCR) |
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== PCR (Nicholas, Ben) 16/1/13== | == PCR (Nicholas, Ben) 16/1/13== | ||
* Repeated PCR on successful samples from November 2012. | |||
* Ran PCR with PB1 primers on these two + 1 positive control with SYBR green Taq readymix + 1 PC with Taq readymix | |||
* 25ul total PCR reaction volume - 5ul template (1ul for PC), 2.5ul each forward & reverse primers, 12.5ul SYBR green Taq readymix / 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. | |||
* PCR left overnight. 7ul of samples loaded, 7ul of ladder loaded. (Less than previous times) | |||
* Ran electropheresis with new, larger power source, with constant 80V. | |||
* After 5 mins: bands were initially visible, especially ladder, Ben, and PC. | |||
* After 40 mins: We left it for too long, and the bands had disappeared. No photo taken. | |||
'''Notes:''' | |||
We could work on a better visualisation system. Currently using black pouch as background. | |||
The gel bath has broken, we will try a plastic food container next time. | |||
The main button on the smaller pipette has broken, super glue needed. | |||
