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PCRboost™ Protocol for enhanced DNA amplification

Product Nos. 63301-001, 62301-021, 63301-011

Product Description

Polymerase chain reaction (PCR) is one of the most performed molecular techniques not only in the academic and industrial research world but also in the forensics, veterinary and health industries. In PCR, DNA is exponentially amplified using Taq DNA polymerase from Thermus aquaticus, a bacterium found in hot springs. PCRboost was designed to enhance the amplification of DNA without the need for time consuming optimizations. PCRboost allows for a decrease in the number of PCR cycles and use of Taq DNA polymerase, resulting in significant time and money savings.

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices. 

Storage/Stability

Store at room temperature. Use within 6 months of purchase date for optimal product performance.

Procedure

When using PCRboost, there is no change to your standard PCR protocol. Simply add PCRboost to your amplification reactions.

The following are examples of PCR and RT-PCR reactions containing PCRboost. Use your own optimized protocol for specific target amplification.

Example PCR reaction (with 500bp amplicon)

Genomic DNA
10x buffer  
dNTP (10mM each)
Primer F (10 µM)
Primer R (10 µM)
DNA Taq Pol
PCRboost
Final volume 
pg to ng
5 µl
1 µl
1 µl
1 µl
2.5-5U
bring up to 50 µl
50 µl


Example PCR Cycling Protocol

94°C
94°C
55-65°C
72°C
72°C
4°C      
2 min
15-30 sec
30 sec
30 sec
4 min
keep


Example RTPCR reactions

First Strand Synthesis
Total RNA
Oligo d(T)
Water
Final Volume

50-100 ng
0.1 µg
bring up to 10 µl
10 µl


Heat at 65°C for 5 minutes. Cool to room temperature for ten minutes, then add:

10x buffer
dNTP (10mM each)
RNAse inhibitor
RT enzyme
2 µl
1 µl
20U
50U


Heat at 42°C for 60 minutes, then 70°C for 15 minutes.

Second Strand Synthesis:

cDNA
10x buffer
dNTP (10mM each)
Primer F (10 µM)
Primer R (10 µM)
DNA Taq Pol
PCRboost
Total
1 µl
5 µl
1 µl
1 µl
1 µl
2.5-5U
bring up to 50 µl
50 µl

Materials

     
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