SNPase "Hot-Start-Polymerase for SNP-Genotyping"

Features:
- 10-15 fold lower mutation rate than Taq DNA Polymerase
- high fidelity allele-specific amplification of DNA fragments
- high specificity with lowest background AS-PEX and AS-PCR
- Hot-Start activity for less primer dimers
- only 5'-3' polymerase activity, lack of 5’-exonuclease activity

Applications:
- High specific PCR
- Single Nucleotide Polymorphism (SNP)
- Multiplex PCR
- Real-Time PCR with intercalation dyes
- high fidelity dNTPs and ddNTPs
- Mini-Sequencing, SNP-genotyping

Description:
SNPase is Taq DNA Polymerase  with unique N-terminal deletion and proprietary amino acids substitutions introduced into the active center of the enzyme. This modification causes dramatic increase of sensitivity of the enzyme to mismatches at 3’-end of the primer. Consequently , non-perfect annealing of the primers does not result in unspecific amplicons formation. This enzyme has only 5'-3' polymerase activity and is recommended for SNP genotyping by allele-specific PCR (AS-PCR), allele-specific primer extension (AS-PEX) and minisequencing procedures.

Minisequencing SNP genotyping with SNPase DNA Polymerase can be carried out by the procedure described in *:

* Reference for minisequencing protocol: Lovmar L, Fredriksson M, Liljedahl U, Sigurdsson S, Syvänen AC.
Quantitative evaluation by minisequencing and microarrays reveals accurate multiplexed SNP genotyping of whole genome amplified DNA. Nucleic Acids Res. 2003;31:e129.

Concentration:
20-25 u/µl

Reaction Buffer supplied:
5X Reaction buffer without MgCl2
MgCl2 100 mM

Note:
- optimal MgCl2 concentration: 3.0 -3.5 mM in the 1X reaction mixture
- higher MgCl2 concentrations results in higher yield (up to 4.5 mM)
- lower MgCl2 (2.5 mM) results in higher specificity
- DNA fragments up to 400 bp from Human genomic DNA and 500 bp from Phage-DNA

Usage:

Components /Volume per reaction
 5X reaction buffer / 5 µl
 MgCl2 / 2.5 - 4 mM
 dNTP-Mix / 0.2 mM each
 primer mix (5 µM stock) / 0,9-1,1 µl (5 pmol)
 Template DNA / 75-125 ng/25 µl genomic DNA
 SNpase (Single Nucleotide Polymorphism) / 0.2 - 0.5 µl  (5-12 units)
 Sterile dest. Water (molecular grade) / up to 25 µl total reaction volume