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Description


Chai’s 2X PCR Master Mix is an optimized ready-to-use solution for routine PCR and qPCR. The mix uses an open Taq polymerase in conjunction with enhancers and stabilizers to deliver exceptional performance at low cost. The Master Mix is compatible with dye and probe based assays, and products between 100 bp and 5 kb can be amplified.

Pcr master mix manual cover @2x

Specifications


Concentration2X
Shipping ConditionsIce
Storage Temperature-20 °C
Exonuclease Activity5' → 3'
Hot StartNo
Recommended Reaction Size10 – 50 µL

Composition


ComponentAmount
Taq polymerase50 U/mL
KCl100 mM
MgCl26 mM
TrisCl pH 8.620 mM
Glycerol10%
Trehalose200 mM
BSA0.4 mg/mL
Detergents0.26%
dNTPs (each)600 µM

Performance


Lambda Phage Standard Curve Amplified with Chai PCR Master Mix 2X
Lambda Phage Standard Curve Amplified with Chai PCR Master Mix 2X
Five ten-fold serial dilutions of lambda phage were used to create a PCR standard curve. A 500 bp product was amplified using Chai PCR Master Mix 2X and Chai Green dye, with an efficiency of 97%, slope of -3.394 and R2 value of 0.996.

Lambda Phage Gel Electrophoresis
Gel electrophoresis image of lambda phage gene fragments amplified with Chai PCR Master Mix 2X
Lambda phage gene fragments amplified using Chai PCR Master Mix 2X. Fragments of 200 bp (lane 1), 500 bp (lane 2), 803 bp (lane 3), 1000 bp (lane 4), and 1400 bp (lane 5) were amplified using lambda phage DNA as template and resolved by agarose gel electrophoresis on a 1% gel.

Protocol


Thaw the 2X PCR Master Mix at room temperature.Vortex the Master Mix gently, and spin it briefly in a micro-centrifuge to collect the material in the bottom of the tube.


Reaction Set Up:

Assemble reaction components on ice and quickly transfer the reactions to a thermocycler preheated to the denaturation temperature (95 ºC). The recommended reaction volume is 25 µL. Reaction volumes between 10 and 50 µL may be used; scale up/down the reaction components accordingly. Final concentration of the Master Mix in the reaction should be 1X.


Component25 µL REACTION VOLUME
10 µM Forward Primer0.5 µL
10 µM Reverse Primer0.5 µL
Template DNA1 ng – 1 µg Genomic
0.5 pg – 5 ng Plasmid / Viral
1 ng – 100 ng cDNA
2X Master Mix12.5 µL
Nuclease-Free WaterBring up to 25 µL

Thermocycling Conditions


Three-Step PCR
StepTempTime
Initial denaturation95 °C1 – 2 min
 Denature95 °C15 – 30 s
 Anneal45 – 68 °C15 – 30 s
 Extend68 °C1 min/kb
Final Extension68 °C5 min
Hold4 °C

  Cycle 30 – 40x

Two-Step PCR

When primer annealing temperatures are above 60 °C, a two-step PCR can be used.

StepTempTime
Initial denaturation95 °C1 – 2 min
 Denature95 °C15 – 30 s
 Anneal/Extension60 °C1 min/kb
Final Extension68 °C5 min
Hold4 °C

  Cycle 30 – 40x

Downloads


Product Manual Download
Safety Data Sheet (SDS) Download

Certificate of Analysis


Lot 4181021 Download