• Instant HiFi PCR MasterMix
  • Instant HiFi PCR MasterMix

Instant HiFi PCR MasterMix

This product is a ready to use 2 × High fidelity PCR pre prepared reaction solution, containing all PCR reagents except primers and templates, including Pfu DNA polymerase, dNTPs, MgCl2, reaction buffer, PCR enhancer, sample dye, etc., is particularly suitable for high fidelity PCR reactions.
$45.00
  • Instant HiFi PCR MasterMix

SPECIFICATION

                          Instant HiFi PCR MasterMix                                                                     
This product is a ready to use 2 × High fidelity PCR pre prepared reaction solution, containing all PCR reagents except primers and templates, including Pfu DNA polymerase, dNTPs, MgCl2, reaction buffer, PCR enhancer, sample dye, etc., is particularly suitable for high fidelity PCR reactions.
Product features:
1. Pfu DNA polymerase is the most widely used high-fidelity enzyme, with a fidelity 10 times that of ordinary Taq enzymes.
2. When in use, PCR reaction can be carried out by simply adding templates and primers and diluting them to 1 concentration, which is very convenient and simplifies the preparation work for PCR.
3. All necessary reagents for PCR reaction are collected in one tube, and the reaction system configuration can be completed in a few minutes. Due to the limited sampling process, it is beneficial to reduce pollution and experimental errors.
4. Type A of this product contains electrophoretic dyes, and the PCR reaction solution can be directly applied for electrophoresis, further simplifying the operation.
5. Due to careful optimization of the concentration and proportion of each component, the reaction has high sensitivity and specificity.
6. Suitable for high-fidelity amplification of genes and gene site-specific mutagenesis experiments, the PCR products obtained can be used for flat end cloning.
Storage conditions: transported at low temperature, stored at -20 ℃, with a shelf life of 12 months.
Usage:
Mix this product with the user provided template and primer according to the following recommended dosage (30 μ L reaction system) can be added to the PCR tube for PCR, and after the reaction is completed, 5-15 can be directly taken μ L electrophoresis amplification results. If the reaction system is not 30 μ L. Each component needs to be increased or decreased proportionally.
Reagent dosage
two × High fidelity PCR MagicMix 25 μ L
DNA template (self prepared) Mammalian genome DNA 0.5-1 μ G
Yeast Genomic DNA 5-500ng
Bacterial genomic DNA 0.5-50ng
Plasmid DNA 5-500ng
PCR recovered fragments 1-100pg
PCR primer (self prepared) 10-20pmol each
Replenish water to 30 μ L
Reference amplification conditions:
Pre denaturation at 94 ℃ for 4 minutes
PCR (30 cycles) 94 ℃ for 30s
50 ℃ for 30s
68 ℃ 0.5kb/min
Extend 68 ℃ for 10 minutes
Precautions:
1. The extension reaction rate of Pfu DNA polymerase in this product is lower than that of Taq DNA polymerase, so our recommended amplification extension reaction time is 2 minutes per kb.
2. The activity of Pfu enzyme 3 '-5' exonuclease can degrade primers, so it is strongly recommended to prepare the PCR reaction solution on ice and immediately place it in a PCR instrument for amplification. After amplification, electrophoresis detection should be carried out immediately.
3. The amplified product of this product is a flat end and cannot be directly cloned into TA. Instead, it can be cloned using the ClionB vector (BTN51104). If TA cloning is required, it is recommended to purify the PCR product and add A with the A reagent kit (BTN60105) before performing TA cloning
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