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Project:Blood typing: Difference between revisions
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* 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== | ==Extraction and PCR (JJ, Bene, Will) 20-23/2/13== | ||
* 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. | * 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. | ||
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[[File:blood-test-will.JPG|left|400px|thumb|none|Will blood test]] | [[File:blood-test-will.JPG|left|400px|thumb|none|Will blood test]] | ||
We bought 5 home testing kits to check our results. Will is A-, as shown here. Each circle contains different types of antigens, and if your blood agglutinates in a circle it means you have that antigen. | We bought 5 home testing kits to check our results. Will is A-, as shown here. Each circle contains different types of antigens, and if your blood agglutinates in a circle it means you have that antigen. | ||
==Extraction and PCR (Simon, Lui, Will) 26/2/13== | |||
* Ran PCR with PB1 primers on JJ's sample from 20/2 and Bene's sample from 23/2. | |||
* Did two reactions for each gDNA sample. 25ul total PCR reaction volume - 5ul dH20, 10ul template, 2.5ul each forward & reverse primers, 12.5ul Taq readymix | |||
* Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | |||
* Ran samples on gel to check PCR succeeded. | |||
===Restriction digest=== | |||
* Incubated JJ's PCR product with Kpn1 at 37C for 65 mins. Restriction digest mix contained 25ul PCR product, 1ul Kpn1, 15ul buffer. | |||
