"GRGGTA" at PICR.CR.MAN.AC.UK
Mon Jan 24 06:08:08 EST 1994
Try Linearising the plasmid, do not phosphatase, phenol extract.
Anneal oligo's in 100mM NaCl (80 degrees C down to RT, then leave on
ice), Do not kinase.
Ligate 25-50ng vector with 1 microgramme annealed oligo 2-4 hrs RT.
Works every time for me (greater than 90% contain inserts) with added
bonus that there are no multiple inserts.
Why solve problems if you can avoid them!
GRGGTA at UK.AC.MAN.CR.PICR
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