5. 2 x Fast High Fidelity PCR Master Mix, 5 mL $300,

Product Information

Description/Introduction

This product is a ready-to-use PCR premix, containing modified ultra-high fidelity DNA polymerase, dNTPs and optimized PCR reaction buffer, with a concentration of 2×. During PCR amplification, only template, primer and ddH2O were added to make the concentration of mix solution 1× The PCR reaction can be performed. The amplification ability is strong, the amplification speed is fast, and the extension speed can reach 5-15 s / KB. It has ultra-high fidelity. The fidelity is 50 times that of Taq DNA polymerase and 6 times that of PFU DNA polymerase. The amplification product is flat end. Loading dye has been added to the product, and the amplified product can be directly used for agarose gel electrophoresis detection. This product is suitable for PCR amplification of conventional PCR, colony PCR, complex template and high GC content template.

Storage and Handling Conditions

Transport with wet ice. Store at -20°C, valid for 12 months.

Component

Assay Protocol / Procedures

1.Recommended PCR reaction system:

a: When using plasmid or phage DNA as template, the recommended dosage is 10ng-1pg/50 μL. When using genomic DNA as template, the recommended dosage is 250ng to 50 ng/50μL. When using cDNA as a template, it is recommended to dilute 2-100 times and add no more than 10% of the total system. Excessive templates can easily lead to non-specific amplification, and too few templates can easily lead to low PCR amplification efficiency;

b: The final primer concentration ranges from 0.2-1.0μM, and the recommended primer concentration is 0.4μM. Too few primers may lead to low yield or no amplification, and too many primers may lead to non-specific amplification;

c: High GC templates can add an additional DMSO up to 10% of the total volume to the reaction system.

2.Recommended PCR amplification conditions:

a: The pre-denaturation time of 30 s is suitable for most conventional templates, and the denaturation time of complex templates can be extended to 2 min;

b: For complex templates, the annealing time can be extended to 30 s for amplification with merge primers

c: The recommended extension speed of plasmid and other simple templates is 5-10 s / KB; The recommended extension speed of conventional genomic template is 10-15 s / KB; Complex template 15-30 s / KB

Note:

1.Thoroughly thaw and mix before use. After complete thawing, it can be kept at 4 ℃ for at least 2 weeks to avoid repeated freezing and thawing.

2.Please wear disposable gloves during operation.

For Research Use Only!

Ver. No.: V1.0-202208