Human Genomic DNA Quantitative Kit
2024-10-25Y-chromosome Human DNA Quantitative Kit
2024-10-25Product Number: PCK84
Shipping and Storage
-20 ℃, 12 months. If frequently used, it can be stored at 2-8 ℃ to avoid repeated freeze-thaw as much as possible.
Components
| Component | PCK84 | PCK84 |
| 2×SYBR qPCR Master Mix | 1 ml | 5×1 ml |
| qPCR Primer Mix | 100 µl | 5×100 µl |
| DNA Standard 1 | 100 µl | 5×100 µl |
| DNA Standard 2 | 100 µl | 5×100 µl |
| DNA Standard 3 | 100 µl | 5×100 µl |
| DNA Standard 4 | 100 µl | 5×100 µl |
| DNA Standard 5 | 100 µl | 5×100 µl |
| 50×High ROX | 40 µl | 200 µl |
Description
This product uses the dye method (SYBR Green I) to perform real-time fluorescence quantitative PCR (qPCR) on the products after NGS library construction. The reagent kit provides the reaction mixture, DNA primer mixture, standards, and complete reagent system required for the qPCR process, making the operation simple and convenient.This kit uses a chemically modified new highly efficient hot start polymerase, and the activation of the enzyme requires incubation at 95℃ for 10 minutes. This product has strong specificity and high amplification efficiency, and can quickly and accurately quantify the concentration of the constructed library. Suitable for fluorescence quantitative PCR instruments that do not require ROX as a calibration dye, such as Roche LightCyler 480, Roche LightCyler 96, Bio radioCyleriQ, iQ5, CFX96.
ROX dye is used to correct the fluorescence signal error generated between wells in quantitative PCR instruments, and is generally used in Real Time PCR amplification instruments from companies such as ABI and Stratagene. The excitation optical systems of different instruments vary, so the concentration of ROX dye must be matched with the corresponding fluorescence quantitative PCR instrument.
Instruments that do not require ROX calibration:Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96 etc.
Instruments that require Low ROX calibration: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 that require High ROX calibration:ABI Prism 7000/7300/7700/7900，Eppendorf，ABI StepOne/Step One Plus etc.
Note:The preparation methods for High Rox and Low Rox are described in protocol Step 2.
Application
This product is designed for the Illumina platform second-generation sequencing library concentration absolute quantification. The end of the library contains Illumina P5 and P7 chip binding sequences, with a length of no more than 1 kb and a concentration of no less than 0.002 pM. This product can be used for quantitative experiments. The qPCR Primer Mix provided by the kit contains the following two primer sequences:
- Primer 1: 5'-AAT GAT ACG GCG ACC ACC GA-3'
- Primer 2: 5'-CAA GCA GAA GAC GGC ATA CGA-3'
It is possible to confirm in advance whether the library can be amplified by the primer pair through the primer sequence.
