2×SYBR qPCR Mix
This product is a ready-to-use PCR kit based on fluorescent dye detection, which can perform real-time quantitative PCR analysis (qPCR) on target fragments. The product contains all the components required for real-time quantitative PCR, including optimized buffer components, Taq DNA polymerase, dNTPs, MgCl2, and fluorescent dyes. Users only need to add templates and primers to perform real-time quantitative PCR reaction and HRM analysis, which has a wide range of applications.
Convenient, users only need to prepare templates and primers for real-time quantitative PCR experiments.
The use of third-generation saturated fluorescent dyes has a strong signal and will not inhibit PCR reactions.
The fast and easy-to-use pre mixed solution greatly simplifies the sampling operation steps, avoids cross contamination, and reduces experimental errors.
The concentration and proportion of each component have been carefully optimized, resulting in high sensitivity and specificity of the reaction. Target molecules with a concentration of 50 copies/mL can be detected.
Component specifications
two × SYBR qPCR Mix 1mL
Storage: -20 ℃, valid for 6 months.
Configuration reaction system
With 20 μ For example, in the qPCR reaction system of L, the following components are added to a clean PCR tube:
Reaction system components 20 μ Final concentration of L system
two × SYBR qPCR Mix 10 μ L 1 ×
Self prepared primer - a x μ L 0.1-0.5 μ M
Self prepared primer II a x μ L 0.1-0.5 μ M
Self prepared template a x μ L
Self provided ROXb 2 μ Select according to different instruments
Supplement ultrapure water to 20 μ L
Place it in a fluorescence PCR instrument for amplification and record the fluorescence signal during annealing or extension steps.
Attention:
Typically, the primer concentration is 0.2 μ M can obtain good results, with a final concentration of 0.1-1.0 μ M serves as a reference for setting the range; Usually, the amount of DNA templates is based on 10-100ng genomic DNA or 1-10ng cDNA. Due to the different copy numbers of target genes contained in templates of different species, gradient dilution can be performed on the templates to determine the optimal template usage.
ROX correction dye:
If using ABI's 75007700 and 7900 models of fluorescent PCR instruments, it is necessary to use the self provided ROX for Ct value correction.
The optimal concentration of ROX for ABI 7700 and 7900 is 1 × (i.e. every 20 μ Adding 2 in the L reaction system μ L 10 × ROX).
The optimal concentration of ROX for ABI 7500 is 0.05-0.1 × (Every 20 μ L reaction system with addition of 0.1~0.2 μ L 10 × ROX; You can also add 10 × Dilute ROX to 1 ×， Then add 1-2 to each reaction system μ L 1 × ROX). ROX will generate noise in the dissolution curve. Therefore, if there are spurious peaks, uncheck the "ROX" option in the "Passive Reference Dye" section of the software and reanalyze the data.
For iCycler IQ, MJ Opticon, MJ Chrono
