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Description


Chai’s Dry PCR Master Mix is an optimized for routine PCR and qPCR. It is lyophilized for ambient temperature shipping and can easily be reconstituted with the provided buffer. 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 dry manual cover

Specifications


Concentration (Reconstituted)2X
Shipping ConditionsAmbient Temp
Storage Temperature-20 – 4 °C
Storage Temperature (Reconstituted)-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


Amplification of Lambda Phage DNA Using Chai PCR Master Mix DRY
Lambda Phage Amplified with Chai PCR Master Mix 2X
Five ten-fold serial dilutions of lambda phage DNA were used as template to amplify a 200 bp product.

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 DRY. 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


Reconstitution:

Remove seal and cap from the vial of lyophilized PCR Master Mix. Add 0.66 mL reconstitution buffer to the freeze-dried vial. Gently pipette up and down at least 10 times to ensure solution homogeneity. Avoid bubbles. Do not vortex. The liquid 2X Master Mix is now ready for use.


Reaction Set Up:

Assemble reaction components on ice and quickly transfer the reaction(s) to a thermocycler preheated to the denaturation temperature (95 °C). 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
Forward Primer0.1 – 0.5 µM
Reverse Primer0.1 – 0.5 µM
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