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badguy

Member Since 03 Feb 2010
Offline Last Active Jun 17 2013 12:20 AM
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Posts I've Made

In Topic: PCR with Plasmid recovered from filter paper

01 April 2013 - 04:48 PM

If that is the case, just elute the plasmid with 50ul of TE buffer or less by suspending and centrifuging at full speed if my memory serves me right.
after that just add in your competent cells and do the transformation.

In Topic: Annealing temperature for PCR

27 March 2013 - 11:28 PM

The formula I used is Ta= 2(A+T)+4(G+C) - 2
I was taught to use that formula and so far so good.

In Topic: PCR with Plasmid recovered from filter paper

27 March 2013 - 11:25 PM

you can use 50ul of TE buffer/nuclease free water to elute the plasmid.
It's better to do transformation rather than doing PCR directly. You never how much of plasmid has been loaded into the paper.
Is the plasmid in FTA card? There are instructions on their website on how to elute plasmid from it.

In Topic: DNA polymerase for GC rich template?

10 January 2013 - 06:49 PM

View PostCurtis, on 09 January 2013 - 07:27 AM, said:

you can't really say. I'm doing some difficult pcr with Finnzyme's Phusion which comes with a GC buffer only for GC rich sequeces. We used Bioline's, it didn't amplify our fragment
What Bioline's polymerase you used?

In Topic: Thermal cycler recommendation

23 December 2012 - 09:48 PM

I have come across this thermal cycler: Speedcycler2, which heating rate of 15C/s and cooing rate of 10C/s.
So far I only see heating rate of 4C/s and cooling rate of 3C/s in most thermal cyclers.

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