Response: Random Hexamer PCR

Robert Slany mfb102 at cd4680fs.rrze.uni-erlangen.de
Wed Nov 3 06:46:59 EST 1993


Dear Lung,

To my opinion the amplification of cDNA with random hexamer primers is of
course not possible. Since you prime on every possible site within your cDNA
(randomly !!) you will get a continous smear of amplification products of 
any possible (random) length. The chance for a correct full length amplification product (requires the binding of one primer out of 4096 possible at both
very outer ends of your cDNA) is 1/4096 * 1/4096 (given that your cDNA
contains all possible 6 mer Sequences, which will  - I admit it- almost never
be the case). But anyway it will be very difficult to purify the correct
product out of the myriad of partial amplifications even on a very high 
resolution gel. 
Hopefully I am wrong now, because this would be a fantastic universal method.
What do you think.
Yours
Robert Slany
-- 

* Robert Slany, Institut fuer Biochemie, Fahrstr. 17, Tel.Ger-09131-854196 *
* slany at imbg.biologie.uni-erlangen.dbp.de oder                             *
* mfb102 at cd4680fs.rrze.uni-erlangen.de                                     *



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