Help Designing Primers

John Dixon jpcd0 at
Mon Jul 25 10:01:19 EST 1994

I need to design primers for sequencing from known sequence using a
DyeDeoxy Terminator Cycle sequencing kit and analysis by an ABI automated
sequencer. So far I have been using MacOligo to pick my primers for me,
but to be honest I don't have a clue how it is selecting them and why it
thinks some are better than others. I realize that you want minimal
dimerization/hairpin and a unique 3'end at least. But can someone tell me
where I can find a good guide to primer design explaining what attributes
a good primer needs for different applications (ie Tm, length, GC richness
etc etc - I dont have MacOligo Manual as we use a communal Networked copy
of the program and I cant find the manual). 

Thanks in advance

John Dixon                     Lab (223) 334131
Wellcome/CRC Institute         Fax (223) 334134
Dept Genetics
Cambridge University           e-mail jpcd0 at
United Kingdom

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