Gene construction advice needed

[Dr. Hiranya S. Roychowdhury]

W/o proper information on how many pairs of oligos, length, RE sites, if
any... it is difficult to talk about a strategy or a protocol for such
projects. 


At 07:59 AM 7/8/00 -0400, Joseph C. Bagshaw wrote:
>I'm looking for advice on a gene construction project.  I'm trying to
>assemble a 219 bp gene from oligonucleotides.  I designed the oligos so
>that each overlaps the next by ten complementary bases.  The plan then is
>to fill the gaps, then ligate.  I can get perfect results with any two
>adjacent oligos, but when I try to assemble the whole, instead of a 219 bp
>band I get a smear.  Any advice or direction to any source of information
>would be greatly appreciated.  I have not found any reference or "how
>to" source, like "Idiot's Guide to Gene Construction".  Please mail to the
>address below or post here.
>
>********************  HAVE GENES, WILL TRAVEL  ********************
>Joe Bagshaw, Worcester Polytechnic Institute
>jbagshaw at wpi.edu
>Roadkill on the information superhighway.
>
>
>
Dr. Hiranya Sankar Roychowdhury
Dept. of Molecular Biology	
PO Box 30001 - 3MLS
New Mexico State University
Las Cruces, NM 88003

Lab: (505) 646 4722
Office: (505) 646 8256
hroychow at nmsu.edu


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