gel

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Shirley
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gel

Post by Shirley » Wed Jan 25, 2006 10:58 am

I have made some competent cells and transform them with a vector. Then I did a plasmid isolation. The DNA concentration is 100ug/ml. I run a gel but no band is detected. So you think my competent cells are really competent?

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canalon
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Post by canalon » Wed Jan 25, 2006 3:23 pm

There are plenty of things that may have gone wrong:
- competent cells as you said,
but also :
- plasmid isolation
- Selection of transformants

Without more details I can't help.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

sdekivit
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Post by sdekivit » Thu Jan 26, 2006 7:20 am

Canalon wrote:There are plenty of things that may have gone wrong:
- competent cells as you said,
but also :
- plasmid isolation
- Selection of transformants

Without more details I can't help.


or the gel isn't made properly

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canalon
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Post by canalon » Thu Jan 26, 2006 3:04 pm

sdekivit wrote:
Canalon wrote:There are plenty of things that may have gone wrong:
- competent cells as you said,
but also :
- plasmid isolation
- Selection of transformants

Without more details I can't help.


or the gel isn't made properly


True enough.
Run in water? (I did that once, it melted :cry: )
Forget to stain?
Did not load? I once had a problem with a loading buffer that wasn't dense enough to make the solution go into the gel. It would just stay on top and leak out :evil: )
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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