927
edits
Project:Blood typing: Difference between revisions
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1) Obtain two sequences of DNA through PCR, the first containing the deletion site at 258, and the second containing the SNP site at 700 | 1) Obtain two sequences of DNA through PCR, the first containing the deletion site at 258, and the second containing the SNP site at 700 | ||
2) Use restriction enzyme ''KpnI'' on the first fragment to cut the O alleles only, and use restriction enzyme ''AluI'' on the second fragment to cut the B alleles only. | 2) Use restriction enzyme ''KpnI'' on the first fragment to cut the O alleles only, and use restriction enzyme ''AluI'' on the second fragment to cut the B alleles only. (KpnI's cutting site is GGTAC^C - in A and B alleles there is a G between the latter two Cs, hence they are not cut. AluI's cutting site is AG^CT - in A and O alleles the initial A is a G, hence only B alleles are cut.) | ||
3) Do gel electrophoresis on the resulting fragments, hopefully resulting in distinguishable bands to show the 6 different alleles. We should then be able to determine an individual's blood type. | 3) Do gel electrophoresis on the resulting fragments, hopefully resulting in distinguishable bands to show the 6 different alleles. We should then be able to determine an individual's blood type. | ||
