Large PCR products

Sebastian W. Bunka seb at i102pc1.vu-wien.ac.at
Thu Feb 3 04:09:22 EST 1994


Jose_Costoya (Jose-Costoya at SEINS.usc.es) wrote:
: Hi, 
:  
:    I would like to get some protocol for amplification of large 
: PCR products (> 1500 bp). 
:  
:         Thanks in advance! 
:                               Jose 
:                           School of Medicine 
:                        University of Santiago de Compostela 
:                               SPAIN 
:                        Jose-Costoya at seins.usc.es 

I've done amplifications of genes of 1700bp using standard protocolls
(25 cycles, 94deg 40 sec, 50 deg 40 sec, 72 deg 1:30 min)

(Mix: 2 ul DNA sol., each 2 ul primer (75 uM), 1 ul dNTP mix (10 mM each),
5 ul 10x polymerase buffer, 38 ul a. dest.; mix, add .5 ul polymerase,
mix, add 50 ul paraffin oil, spin 5 sec., cycle .....)
(you can try also adding BSA; thats not required with several polymerases).

and with the same protocoll fragments of 3250bp; works fine!
(I KNOW :-) the Dynazyme polymerase from Finnzymes Oy works great!
but kkep in mind the patent of LaRoche on the PCR process ....)

Cheers
SWB

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