Solid-state library amplification

Dr. S. Bhattacharya sbhattac at crc.ac.uk
Thu Jan 6 06:05:46 EST 1994


Hi netters

I am writing about a problem I encountered using  the solid-state
library amplification method described by Michael Kriegler (Gene
Transfer and Expression - a lab manual, Stockton Press NY), p 131.

In this method, the bugs (containing plasmid dna), are grown in a gel matrix
(0.3% SeaPrep agarose- LB), the theory being that colonies become nutrient
limited in 3 dimensional space and slower growing colonies catch up with
faster ones. The bugs are then spun out of the gel matrix and maxi-prepped as 
usual.

Brilliant I thought.

All went well until the final step. On spinning the gel matrix containing the 
bugs, at 8000 rpm as recommended, the agarose spun out with the bugs, causing
much despondence.  I was able to melt the agarose and recover the bugs and
library, but what went wrong?

I had used Boehringer agaroseMP -  not having Sea-Prep, could this be the
cause?

I would be grateful for any comments.

Shoumo
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Shoumo Bhattacharya		
MRC Molecular Medicine Group
Royal Postgraduate Medical School
Hammersmith Hospital, LondonW12 0NN UK
Tel:081-7435566 ext.2343
Fax:081-7498341
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