BioDiamond 2X PCR Master Mix Red 1.25ml 混合酵素
Cat No. DMDRT001
Pack size: 4x 1.25 ml
qPCR Instruments:
BioRad CFX96, Roche LightCycler 480, MJ Research Opticon and Opticon 2, MJ Research Chromo 4, Corbett Rotor-
Description
2X Redy Mix is optimized mixture contain of Taq enzyme, reaction buffer, dNTP, enhancer and red dye as 2-fold concentration. 2x Redy mix is designed to allow the user for quick ,easy preparation and ready loading of reaction mixture. The 2x Redy mix can be amplification PCR products up to 3 kb and the products can be directly cloning into T-vector.
Storage conditions
at -20°C
Template
2 x Redy mix is suitable for amplifying targets up to 3 kb from the following templates:
Genomic DNA: 10–200 ng
Plasmid DNA: 1–5 ng
cDNA: ~100 ng starting total RNA
Primers
Use 0.3 μM per primer as a general starting point. For larger amounts of template (e.g., 200 ng genomic DNA), increasing the concentration up to 0.5 μM per primer may improve yield.
Annealing Temperature
The annealing temperature is slightly higher than with typical PCR. The optimal annealing temperature should be ~2°C lower than the Tm of the primers used. A range of 58–68°C is recommended.
Extension Time
As little as 30 seconds per kb is suitable for most targets. Use up to 60 seconds per kb for maximum yield.
Major composition:
NH4 + buffer system with 3.0mM MgCl2
Taq
dNTP mix
PCR Protocol:
1.Thaw the 2x Redy mix at room temperature. Vortex the 2x Redy mix and then spin it briefly in a micro centrifuge to collect the material in the bottom of the tube.
2.Prepare one of the following reaction mixes on ice:
2X Redy Mix is optimized mixture contain of Taq enzyme, reaction buffer, dNTP, enhancer and red dye as 2-fold concentration. 2x Redy mix is designed to allow the user for quick ,easy preparation and ready loading of reaction mixture. The 2x Redy mix can be amplification PCR products up to 3 kb and the products can be directly cloning into T-vector.
Storage conditions
at -20°C
Template
2 x Redy mix is suitable for amplifying targets up to 3 kb from the following templates:
Genomic DNA: 10–200 ng
Plasmid DNA: 1–5 ng
cDNA: ~100 ng starting total RNA
Primers
Use 0.3 μM per primer as a general starting point. For larger amounts of template (e.g., 200 ng genomic DNA), increasing the concentration up to 0.5 μM per primer may improve yield.
Annealing Temperature
The annealing temperature is slightly higher than with typical PCR. The optimal annealing temperature should be ~2°C lower than the Tm of the primers used. A range of 58–68°C is recommended.
Extension Time
As little as 30 seconds per kb is suitable for most targets. Use up to 60 seconds per kb for maximum yield.
Major composition:
NH4 + buffer system with 3.0mM MgCl2
Taq
dNTP mix
PCR Protocol:
1.Thaw the 2x Redy mix at room temperature. Vortex the 2x Redy mix and then spin it briefly in a micro centrifuge to collect the material in the bottom of the tube.
2.Prepare one of the following reaction mixes on ice:
3. If necessary you can scale up your volume
1. Program the thermal cycler as follows:
Step
After cycling, maintain the reaction at 4°C. Samples can be stored at –20°C until use.
Analyze products using standard agarose gel electrophoresis.