Product Name

Squeeze-1-Drop Do PCR Master Mix (with dye, in dripping bottle, No pipetting)

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Application

Squeeze 1 drop from dripping bottle

Specifications

The easiest and most convenient way to run PCR: squeeze then run.

User can squeeze one drop of PCR Master mix into PCR tube/plate, add template and primers, and then can run PCR reaction. This PCR Master Mix contains Taq DNA Polymerase, PCR buffer, Mg2+, dNTP, PCR stabilizer and PCR enhancer. This unique Mix recipe makes the system very reliable, and tolerant to most of the PCR condition (concentration of the components).

The Taq Polymerase in the PCR Master Mix is Hot Start polymerase, which activity is inhibited at ambient temperatures by the chemical modificaiton. This prevents the formation of misprimed products and primer dimers at ambient temperatures. So no need to prepare the reactions on ice. The Polymerase is activated by a 10 minutes, 95°C incubation.

This PCR Master Mix is very stable, and can be stored in 4^oC for up to 6 months. No need to wait the thawing.

The amplification range of the Taq is ~ 6 kb.

The PCR Master Mix contains dye, and can directly run electrophoresis after PCR reaction.

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Manual (protocol)

PCR reaction system

Note: This unique Mix recipe makes the system very reliable, and tolerant to most of the PCR condition (concentration of the components).

Generally, in 25 µL PCR reaction (1 drop volume), the DNA template amount should be 1 pg ~ 10 ng for plasmid DNA template; 1 ng ~ 100 ng for bacterial genomic DNA template; 10 ng ~ 300 ng for eukaryotic genomic DNA template. Too much template DNA may inhibit the PCR reaction.

PCR reaction conditions

Note:

• The recommended annealing temperature is about 5℃ below Tm of primers. If extra bands are observed, higher annealing temperatures should be considered. The absence of product indicates the need for a lower annealing temperature.
• PCR extension time is depended on the size of target gene sequence. Es Taq DNA polymerase is approximately 1 kb DNA / 60 seconds.
• The number of PCR cycles will basically depend on the downstream application of the PCR product.

This PCR Master Mix contains dye for electrophoresis. After PCR, directly load 5 µL of PCR product to agarose gel to run electrophoresis. No need to add loading buffer.