I am interested how can I obtain sufficient quantities of 3 to 16kb DNA fragments from genomic DNA in the fastest possible manner?
use amplitaq gold and do rePCR by making templetes from PCR products itself
- NEB phusion is a highly processive hot start polymerase that might do what you require:
- https://www.neb.com/products/m0530-phusion-high-fidelity-dna-polymerase
- https://www.neb.com/protocols/1/01/01/pcr-protocol-m0530
Takara LA taq is both fast and efficient.
http://www.clontech.com/DK/Products/PCR/Long_PCR/LA_Taq_DNA_Polymerase?sitex=10122:22372:US
Using 2 step PCR can greatly shorten the rxn time.
For example you can use an enzyme like Takara SpeedSTAR (http://www.clontech.com/US/Products/PCR/Fast_PCR/SpeedSTAR_HS_DNA_Polymerase) and use the protocol: 30 cycles of [98℃, 5 sec.; 68℃, 10 sec. (or up to 20 sec.)/kb].Of course you should select the Tm of your primers to be compatible with high annealing temperature, but this can also ensure more specific amplification.
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