HOW to Eliminate PCR primer dimer

Natalie Bloch natalieB at qimr.edu.au
Sun Feb 2 23:55:13 EST 1997


educe primees dimers: add the PRIMERS last and ON ICE in the tubes;
 reduce the quantity of primers;
 the increase in annealing temp will only help reduce the 
unspecific products.
To obtain a stronger signal you could add 1 or 2 cycles, 
or dilute the product obtained 1/100 and rerun another PCR
 with only 25 cycles
Good luck
Natalie.

: Dear netter:
: I have Some questions about PCR (polymerase chain rex) 
:  and need your advices !

: 1.How can I remove (or reduce) the primer dimer in PCR reaction
:    how about increase annealing Temp ? or lowering Ma2+ conc.

: 2.After 38 cycles, a faint signal was obtained. Can I increase 
:    cycle number, or template DNA  or even dNTP content 
:    TO further INCREASE THE SIGNAL I WANT ??

: 3.What's the best methods to elute out the PCR product with
:    high yields and low DNase contaminated. ANY kits suggested ?


: regards    




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