Project:Blood typing: Difference between revisions

Project:Blood typing (view source)

973 bytes added , 24 February 2013

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 * 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]]
+[[File:gel1-23feb2013.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)
+<br style="clear: both" />
+'''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).
+===Extraction and PCR repeat===
+* Bene did a new extraction using same protocol.
+* Ran PCR with PB1 primers on this + JJ's old sample + Will's sample from
+* 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:gel1-23feb2013.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)