SBB11Ntbk-Gary Dixon: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 1: Line 1:
==~~!~~==
==~~!~~==
==[[User:Gary Dixon|Gary Dixon]] 13:22, 17 February 2011 (EST)==
Running two preparative gels:
53/31 PCR = GD1 (#3)
32/53 PCR = GD2 (#4)
5ul of product
5ul of loading dye
And one analytical gel:
42/42= GD3 (#6)
2ul of product
5ul of loading dye
==[[User:Gary Dixon|Gary Dixon]] 14:32, 15 February 2011 (EST)==
==[[User:Gary Dixon|Gary Dixon]] 14:32, 15 February 2011 (EST)==
Performing 3 PCR reactions. One for part sbb1117, and two for part sbb1127 (soeing).
Performing 3 PCR reactions. One for part sbb1117, and two for part sbb1127 (soeing).

Revision as of 11:22, 17 February 2011

~~!~~

Gary Dixon 13:22, 17 February 2011 (EST)

Running two preparative gels:

53/31 PCR = GD1 (#3)

32/53 PCR = GD2 (#4)

5ul of product

5ul of loading dye


And one analytical gel:

42/42= GD3 (#6)

2ul of product

5ul of loading dye

Gary Dixon 14:32, 15 February 2011 (EST)

Performing 3 PCR reactions. One for part sbb1117, and two for part sbb1127 (soeing).

First, sorted out some confusion about letter R and F on oligo. R and F don't necessarily correspond to your forward and reverse oligo. So ss53r is the forward oligo for P_nmpC and ss53f is the reverse.

Then diluted oligos gd031 and gd032.

Then went through the standard procedure to set up PCR reactions. I have three in total.

ss53r/gd031R (718bp, gp = A)

gd032F/ss53f (287bp, gp = B)

ss42r/ss42f (1032bp)

The procedure: http://openwetware.org/wiki/Template:SBB-Protocols_PCR2

Put all three PCR reactions in the <2,000kb reaction.

Gary Dixon 23:59, 5 February 2011 (EST)

Hello World