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Project:Blood typing: Difference between revisions
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* Odd that PC gDNA travelled further in gel than our extractions. | * Odd that PC gDNA travelled further in gel than our extractions. | ||
* Also more confirmation that there's seemingly no correlation between strength of gDNA and strength of PCR product. | * Also more confirmation that there's seemingly no correlation between strength of gDNA and strength of PCR product. | ||
==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. | |||
* 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 | |||
* Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | |||
[[File:gel23feb2013.JPG|left|400px|thumb|none|Gel 23 Feb]] | |||
* 20ul of PCR samples loaded, 20ul of gDNA samples loaded. 4ul loading buffer. | |||
*1: JJ PCR product (no band) | |||
*2: Bene PCR product (very faint band) | |||
*3: PC PRC product (band) | |||
*4: JJ gDNA (band) | |||
*5: Bene gDNA (band) | |||
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'''Notes/Conclusions:''' | |||
* PCRs either failed or were too weak to show. Differences in this reaction that could have affected the result were: we doubled PCR quantities, then loaded 20ul from the result. gDNA bands were fine, so strange PCR didn't work as PC was ok (but not strong). | |||
