2×Long-HiFi PCR MasterMix
Long fragment DNA fast high fidelity PCR MasterMix (with red tracer dye) containing Long HiFi DNA Polymerase, dNTPs, and optimized reaction buffer at a concentration of 2 ×。 When using, simply add templates, primers, and replenish water until the final concentration of Mix is 1 × That's it. Long HiFi DNA Polymerase is a mixture of various ultra fidelity enzymes modified using state-of-the-art genetic engineering with extension factors, greatly improving amplification length, amplification speed, fidelity, and yield. Long HiFi PCR MasterMix is the preferred product for long segment ultra fidelity rapid amplification. This product contains a red tracer dye and can be directly loaded for electrophoresis without the need for sample loading buffer; It can also be purified for subsequent operations such as enzyme digestion, ligation, and fluorescence sequencing.
Fast amplification speed: The extension speed can reach 3-4 kb/min, which is 6-8 times that of pfu.
High amplification yield: Generally, the PCR product yield is 50% to 100% higher than that of traditional PFU.
Excellent fidelity: The fidelity is more than 54 times that of TAQ. Generally, randomly selecting a bacterial colony for sequencing is the correct mutation free colony.
Long amplification length: Using complex genomic DNA as a template is suitable for amplification of products that do not exceed about 10kb, while using simple genomic, plasmid, and phage DNA as templates is suitable for amplification of products that do not exceed about 15kb.
Component 1mL 5mL
two × Long HiFi PCR MasterMix 1mL 5mL
1 instruction manual and 1 copy
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Long HiFi DNA Polymerase amplification speed is fast, and simple templates such as plasmids and simple genomes can be used in 10-15 seconds/kb; Complex templates such as the human genome can be used at 20-25 seconds per kb.
If there is a blurry tail dragging phenomenon on the main band during electrophoresis, or if there is a smear band dragging down from the sample hole in the swimming lane, it generally indicates that the extension time is too long
Long, gradient shortened extension time (recommended 10-15 seconds/kb for gradient reduced extension time) until satisfactory results are obtained.
For templates with high GC content, the pre denaturation and denaturation temperatures can be increased to 98 ℃. Long HiFi DNA Polymerase has strong heat resistance, and the activity of Long HiFi DNA Polymerase remains unchanged at 98 ℃.
If the GC content of the amplification template is high or the template is complex and the amplification effect is poor, DMSO can be added to the reaction mixture to the final concentration of 1% to 8%, and the optimal concentration can be explored by increasing it according to a 1% gradient. Alternatively, add betaine to the final concentration of 1.0-1.7M. And use Touchdown PCR.
Suggested PCR system:
Composition 25 μ L system 50 μ Final concentration of L system
two × Long HiFi PCR MasterMix 12.5 μ L 25 μ L 1 ×
Forward Prim
