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Project:Blood typing: Difference between revisions
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* 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. | ||
[[File: | [[File:gel10jan2013.JPG|left|400px|thumb|none|Gel 10 Jan]] | ||
* 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: 100-3000bp ladder. Ladder [http://www.nbsbio.co.uk/product.asp?pID=8951&cID=33 key]. | *1: 100-3000bp ladder. Ladder [http://www.nbsbio.co.uk/product.asp?pID=8951&cID=33 key]. | ||
*2: Jim PCR product (No band) | *2: Jim PCR product (No band) | ||
*3: Mike PCR product (No band) | *3: Mike PCR product (No band. Blob is result of accidentally touching gel with dirty pipette) | ||
*4: Will PCR product (No band) | *4: Will PCR product (No band) | ||
*5: PC PCR (band) | *5: PC PCR (band) | ||
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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. | |||
