SuperStar Probe Mixture
2023-07-14
Multiplex PCR MasterMix (UNG)
2023-07-17
Cat:PCM63
Storage Condition
-20°C. For frequent use, it can be stored at 2-8°C. Try to avoid repeated freezing and thawing.
Product Component
| Component | PCM63 | PCM63L | PCM63H |
| 2×HotStar Probe Mix UNG | 5×1 ml | 5×1 ml | 5×1 ml |
| 50×Low ROX | - | 200 μl | - |
| 50×High ROX | - | - | 200 μl |
| ddH2O | 5×1 ml | 5×1 ml | 5×1 ml |
Description
2×HotStar Probe Mix UNG is a premix system dedicated to real-time fluorescent quantitative PCR with a probe method (TaqMan, Molecular Beacon, etc.), with a concentration of 2×, including HotStar Taq DNA polymerase, PCR Buffer, dNTPs (dTTP is all replaced by dUTP replaced), UNG enzyme and Mg2+, the operation is simple and convenient. It is mainly used for the detection of genomic DNA target sequences and cDNA target sequences after RNA reverse transcription, such as gene expression analysis, copy number analysis, SNP genotype analysis, etc. The dUTP-UNG anti-pollution system is used in this product, and dUTP is added in the preparation process of the PCR reaction system, so an amplification product containing dU bases is formed. This product can be eliminated by UNG enzyme treatment in the PCR system before the next PCR reaction. In this way, the residual contamination of the PCR product is effectively removed, and the false positive caused by the contamination of the amplification product is greatly reduced. The UNG enzyme can be inactivated by the pre-denaturation step in the PCR cycle, so it will not affect the formation of new dU-containing PCR products. The HotStar Taq DNA Polymerase contained in this product is a chemically modified, brand-new high-efficiency hot-start enzyme, which has no polymerase activity at room temperature, and effectively avoids non-specific binding of primers and templates or primer dimers at room temperature. To generate non-specific amplification, enzyme activation must be incubated at 95°C for 10 minutes. The unique combination of PCR buffer system and hot-start enzyme significantly improves the amplification efficiency of PCR, with stronger fluorescent signal and higher sensitivity, and can detect single-copy templates. Using this product can get a wider linear range and more accurate quantification of the target gene.
ROX dye is used to correct the fluorescence signal error between the wells of the quantitative PCR instrument, and is generally used in Real Time PCR amplification instruments of companies such as ABI and Stratagene. The excitation optics of different instruments are different, so the concentration of ROX dye must be matched with the corresponding real-time PCR instrument.
Instruments that do not require ROX calibration (PCM63):
Roche LightCycler 480, Roche LightCyler 96, Bio-radi iCyler iQ, iQ5, CFX96, etc.
Instruments requiring Low ROX calibration (PCM63L):
ABI Prism7500/7500 Fast, QuantStudio® 3 System, QuantStudio® 5 System, QuantStudio® 6 Flex System, QuantStudio® 7 Flex System, ViiA 7 system, Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000 etc.
Instruments requiring High ROX calibration (PCM63H):
ABI Prism7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus etc.
Note
1. Before use, please mix it upside down gently, try to avoid foaming, and use it after short centrifugation.
2. Avoid repeated freezing and thawing of this product, as repeated freezing and thawing may reduce product performance. Long-term storage of this product can be stored at -20°C, away from light. If it needs to be used frequently in a short period of time, it can be stored at 2-8°C.
