High-Taq DNA Polymerase with 10mM dNTP Mix (4x250U): High-Taq DNA Polymerase with 10mM dNTP Mix (5000U). High-Taq DNA Polymerase (2.5U/ I). OX High-Taq reaction Buffer. 10 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
High-Taq DNA Polymerase (2.5U/ I)
OX High-Taq reaction Buffer
10 mM dNTP Mix (each 1OmM}
• lOX High-Taq reaction Buffer
• 10 mM dNTP Mix (each 1OmM}
• Band Sharpener
A. Template (temperature cycle}
.A imal genomic DNA
SD-200 ng (3D-45 cycles) 1D-50 ng (35-50 cycles)
.B cterial genomic DNA
1D-50 ng (30-40 cycles) 1-5 ng (35-50 cycles)
.Plasmid and lamda DNA
1-5 ng (35-•m cycles)
B. High-Taq DNA Polymerase
1.25unit(0.51JI) per 501JI 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 condHions. 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 (5-201JI for 501JI 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 beG or C.
– It is recommended that Tm of the designed primers is >64’t and AT >58’t.
E. Extension time
– Normally, extension time is Imin/kb. In case target is >3kb, the extension time should be increased to 1.5-2.0 min/kb.
