Stuart McKellar: /* Entry title */

Stuart McKellar — Tue, 11 Dec 2012 02:53:58 GMT

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-	==~~Entry title~~==	+	==PCR with positive controls==
-	* ~~Insert content here~~...	+
		+	Aim: See if we can identify the positive controls using both primers.
		+
		+	Materials and methods: We will be running 5 x PCR reactions: male and female with p2/p8, male and female with 2550/2178 primers, and a negative control. We will run it at 60°C.
		+
		+	reaction mix:
		+	*2.5ul F primer
		+	*2.5ul R primer
		+	*2.0ul dNTPs
		+	*0.5u Taq
		+	*1ul DNA
		+	*2.5ul 10x buffer
		+	*1.5ul MgCl<sub>2</sub>
		+	*13ul H<sub>2</sub>O
		+
		+	Master Mix:
		+	12.5 F
		+	12.5 R
		+	0.5 Taq
		+	10 dNTPs
		+	12.5 Buffer
		+	7.5 MgCl<sub>2</sub>
		+	65ul H<sub>2</sub>O
		+
		+	Negative control used p2/p8 primers as they are the most likely ones to work at these temps. Primers were not added to MM, but instead to each individual tube.
		+
		+	repeated experiment at 58°C.
		+
		+	ran PCR for 35 cycles, annealing temperature is 60°C
		+
		+	looks like faint bands are appearing. I may need to alter the times a little bit, maybe extend the amount of time for elongation. Two bands were distinguishable on the 58 and 60C PCRs when viewed on a 1% gel.
		+
		+	The second one which was 58C annealing temp ran on a 1% gel. I want to try 2550 primer set at 55C today. I might run the 2550 samples on the tawny's and the eclectus.
		+
		+	***DO SOME READING OF THE PAPERS AFTER PCR HAS STARTED***
		+
		+

Stuart McKellar: Autocreate 2012/12/10 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing

Stuart McKellar — Mon, 10 Dec 2012 01:14:26 GMT

Autocreate 2012/12/10 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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==Entry title==
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Stuart McKellar: Autocreate 2012/12/10 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing