This product is designed for PCR amplification of genomic DNA containing uracil after heavy sulfite modification. When genomic DNA is treated with bisulfite, unmethylated cytosine (C) is converted to uracil (U), while methylated cytosine (5-mC or 5-hmC) is not converted, and the methylation of the DNA can be detected by PCR amplification or sequencing. The performance of PCR reaction may be affected by the DNA modified by bisulfite, so the DNA Polymerase and the reaction buffer are optimized to make them suitable for the amplification of genomic DNA modified by bisulfite. For the target fragments that are difficult to amplify, the reaction buffer can be optimized by adjusting Mg2+ or dNTP concentration to regulate amplification efficiency and reaction specificity.
A monoclonal antibody that inhibits the activity of DNA polymerase at room temperature is added to this product, enabling Hot Start PCR, which effectively inhibits the formation of primer dimers and non-specific amplification. The antibody is denatured in the initial DNA denaturation step of the PCR reaction and can be used under normal PCR reaction conditions.
① The performance of PCR reaction may be affected by the DNA modified by bisulfite, so this product optimizes the DNA Polymerase and the reaction buffer to make them suitable for the amplification of genomic DNA modified by bisulfite.
② This product contains all the necessary reagents for the PCR reaction, and each component is individually packaged for easy adjustment of MgCl2 and the reaction concentration of dNTPs to obtain the best experimental results.
③ A monoclonal antibody that inhibits the activity of Taq enzyme is added to this product, which can be used for hot start reaction. The antibody binds to the enzyme and inhibits Taq enzyme activity before the PCR reaction, effectively inhibiting the formation of primer dimers and non-specific amplification.
| individual parts making up a compound | norm |
| MethyTect Taq HS DNA Polymerase (5 U/μl) | 50 μl |
| 10X MethyTect PCR Buffer (Mg)2+ free) | 1 ml |
| dNTP Mix (10 mM each) | 300 μl |
| MgCl2 Solution (50 mM) | 600 μl |
Transportation temperature: dry ice or -20℃ ice pack transportation
