High-Taq DNA Polymerase with 10mM dNTP Mix (250U): High-Taq DNA Polymerase with 10mM dNTP Mix (5000U)
High-Taq DNA Polymerase (2.5U/ I)
OX High-Taq reaction Buffer
10 mM dNTP Mix (each 1OmM}
Band Sharpener
A. Template (temperature cycle)
– Animal genomic DNA
50-200 ng (30-45 cycles)
10-50 ng (35-50 cycles)
– Bacterial genomic DNA
10-50 ng (30-40 cycles)
1-5 ng (35-50 cycles)
– Plasmid and lamda DNA
1-5 ng (35-40 cycles)
B. High-Taq DNA Polymerase
1.25unit (0.5µI) per 50µI reaction is recommended when amplifying animal genomic DNA.
Please use 2.5-5 unit in case target size is >3kb.
In case the target is >3kb, we recommend EF-.Taq
C. Band Sharpener
Band Sharpener Is not necessary for regular PCR conditions. In case fragment include high GC region or hard to amplify
complex secondary structures, please add Band Sharpener to a final concentration of 0.5x-2x (5-20µI for 50µI reaction) to reaction mixture, as optimization is required (see protocol below).
D. Primer design
– Primer can be designed using a primer design software or manually.
– Avoid repeated sequence at 3′ end.
– In case 3′-end is G+C rich, the end have to be A or T.
– In case 3′ end is A+T rich, the end have to be G or C.
– It is recommended that Tm of the designed primers is >64C and AT >58C.
E. Extension time
– Normally, extension time is 1 min/kb. In case target is >3kb, the extension time should be increased to 1.5-2.0 min/kb.
