Page 21 - 2022-23 BIOFACT Catalogue

DNA / RNA Amplification Description

Hot Start PCR Description

Most of the DNA polymerases can extend misaligned primers to create primer-dimers or non-specific product before reaching
the denaturation temperature that has been set. Hot start PCR is a modi ed technique that is developed to avoid non-specific
amplification and limit primer-dimer formation during reaction preparation. By blocking polymerase activity prior to PCR cycling,
non-specific amplification can be reduced and target yield is increased. A variety of Hot start methods exist including chemical
modifications, wax barrier methods and inhibition by a Taq-directed antibody.
★ It is less likely to occur non-specific annealing while rising up to 95℃ because Hot start Taq is activated only at ★ temperature.

Denaturation
It is recommended to use Hot Start PCR method in the following cases :
• High specific PCR 95℃ 95℃ Extension
72℃
• PCR difficulty due to high GC content or secondary structure 55℃ 55℃ 72℃
• PCR difficulty due to non-specific product or primer-dimer
Annealing
• Multiplex PCR / Allele specific PCR

BIOFACT Hot Start PCR Enzymes
Chemical-mediated Hot Start Antibody-mediated Hot Start
- High specificity for targeted genes - Reliable ampli cation for template DNA containing low concentrations
- Pre-denaturation time : 2 ~ 15 mins. - Pre-denaturation time : 2 min.
BioFACT™ F-Star Taq BioFACT™ A-Star Taq DNA Polymerase
BioFACT™ H-Star Taq BioFACT™ A-Star Lamp Taq DNA Polymerase
chemical
SimpleWay™ Direct PCR
5’ 3’
BioFACT™ 2X Multi-Star PCR Series Heating &
PCR
BioFACT™ OneStep RT-PCR
3’ 5’
BioFACT™ Real-Time PCR Series
Chemical-mediated
Target DNA BioFACT TM H-Star Taq Primer dNTP
Chemical-mediated Hot Start Antibody-mediated Hot Start
chemical Anti-Taq antibody + Taq
5’ 3’ 3’
Heating & Heating &
PCR PCR
3’ 5’
3’ 5’
Chemical-mediated
Target DNA Primer dNTP Target DNA Anti-Taq antibody + Taq Primer dNTP
BioFACT TM H-Star Taq
Anti-Taq antibody + Taq
BIOFACT Product
3’
H-Star Taq DNA Polymerase (p. 84), A-StarTaq DNA Polymerase (p. 86), A-Star Lamp Taq DNA Polymerase (p. 90)
Heating &
PCR
2X Multi-Star PCR Series (p. 96)
3’ 5’
Target DNA Anti-Taq antibody + Taq Primer dNTP
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BIOFACT CATALOGUE