pTopo purification problems

Taliaferro, Dwayne (NIH/NIMH) [F] via methods%40net.bio.net (by taliaferrod from mail.nih.gov)
Thu Sep 10 16:31:20 EST 2009


Um...try it again?

On a constructive note...

What size is the gene?
Did you -tail it?
Did you try a control reaction?
Perhaps it was a plasmid prep issue?
Are you sure the colony you picked was bacteria vs fungus?
Are you sure your AMP is working?

On 9/10/09 3:54 PM, "Yoram Gerchman" <gerchman from research.haifa.ac.il> wrote:



Greetings netters
I have recently made an attempt to clone a short gene into pTopo (using Top10
competent cells). i did get white colonies on LB-Amp-X-gal, and they grew
nicely on liquid LB-Amp, but after plasmid purification I get no DNA (tried
already 2 different kits). Any ideas as to how remediate this situation?
Many thanks
Yoram



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