Hot-Start Gene RED PCR Mix

Hot-Start Gene RED PCR Mix is a 2× premixed PCR reagent that converts the conventional Gene RED PCR Mix Plus into a hot-start format using an antibody-mediated method. PCR can be performed simply by adding template DNA and primers.

Product features

• A ready-to-use, two-color premix reagent for quick and easy PCR.
  Simply add template DNA and primers — the reaction mixture can be loaded directly onto an agarose gel after PCR.
• The Hot Start PCR method prevents non-specific amplification using an Anti-Taq antibody.
• Provides high-yield amplification optimized for fast PCR (10 sec/kb).
• Suitable for GC-rich (up to 69%) and long DNA templates (up to 10 kb).

Applications

• Colony PCR
• Insert checking
• TA cloning

PCR performed with primers that tend to produce non-specific amplification

A region of approximately 600 bp was amplified using a primer that often causes non-specific amplification. PCR was performed with annealing temperatures of 65°C, 68°C, 69°C, and 72°C.

M: OneSTEP Marker 5
1: Annealing temperature 65℃
2: Annealing temperature 68℃
3: Annealing temperature 69℃
4: Annealing temperature 72℃
In 2% Agarose S/TAE gel

Non-specific amplification was suppressed at all tested annealing temperatures when using the Hot-Start Gene PCR Mix compared with Gene RED PCR Mix Plus.

PCR performed with primers that tend to form primer dimers

A region of approximately 6 kbp was amplified using a primer that often causes non-specific amplification due to primer dimer formation.PCR was performed with 5 ng, 1 ng, and 500 pg of template DNA.

M: OneSTEP Marker 6
1: 5 ng
2: 1 ng
3: 500 pg
In 1% Agarose S/TAE gel

Primer dimer formation was suppressed, and target region amplification was increased when using the Hot-Start Gene PCR Mix compared with Gene RED PCR Mix Plus.