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제품 설명
4X CAPITAL™ 1-Step qRT-PCR Green Master Mix
제품 번호
BR0502301 - 200 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) 200 µl RTase with RNase Inhibitor
BR0502302 - 1000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) 200 µl RTase with RNase Inhibitor
BR0502303 - 4000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) 200 µl RTase with RNase Inhibitor
BR0502401 - 200 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) LROX 200 µl RTase with RNase Inhibitor
BR0502402 - 1000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) LROX 200 µl RTase with RNase Inhibitor
BR0502403 - 4000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) LROX 200 µl RTase with RNase Inhibitor
BR0502401 - 200 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) HROX 200 µl RTase with RNase Inhibitor
BR0502402 - 1000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) HROX 200 µl RTase with RNase Inhibitor
BR0502403 - 4000 reactions of 20 µl / 1 ml CAPITAL qPCR Green Master Mix (1step) HROX 200 µl RTase with RNase Inhibitor
제품 특징
Description
biotechrabbit CAPITAL™ qRT-PCR Green Mix allows sensitive and specific cDNA synthesis and qPCR in a single tube for quantifying mRNA, total RNA and viral RNA sequences. Extremely low-copy-number targets can be detected with high efficiency over several logs of template concentration.
CAPITAL qRT-PCR Green Mix uses proprietary reverse transcriptase technology and buffer chemistry for efficient cDNA synthesis and QPCR in a single tube. To enable the use of the kit on qPCR platforms with different reference dye concentration requirements, three kit formats are available: a one-step kit containing no ROX, as well as LRox and HRox versions containing ROX in the corresponding concentratios.
Info: Recommended annealing temperature is 2°C above primer Tm (use gradient PCR to optimize the annealing temperature).
Other Information
Quality ControlFunctional assayMix tested functionally in QRT-PCR. |
Protocols
RNase contamination is an exceptional concern when working with RNA. RNase A, providing most threat to RNA integrity, is a highly stable contaminant of any laboratory. To prevent RNA from degradation and to minimize possibility of contamination One Step RT-PCR; follow the guidelines below:
Component | Volume | Final concentration |
Primer Mix (Reverse and Forward) | Variable | 100–400 nM |
Too high primer concentrations result in unspecific amplification and should be avoided. | ||
Template RNA | Variable | 0.01 pg to 1 µg |
Use 1 pg – 1 µg Total RNA, or >0.01 pg mRNA | ||
CAPITAL qPCR Green Mix (1step), 4× | 5 µl | 1× |
RTase with RNase Inhibitor, 20× | 1 µl | 1× |
Nuclease free water | Variable |
|
Total volume | 20 µl |
|
Step | Temperature | Time | Cycles |
Reverse Transcription | 50°C | 10 min | 1 |
Initial activation | 95°C | 3 min | 1 |
Denaturation | 95°C | 10 s | 40-45 |
Annealing/Extension* | (60-68°C) | 30 s |
*Recommended annealing/extension temperature is primer Tm +2°C. Use gradient PCR to optimize the annealing temperature. Do not use temperatures below 60°C. Do not exceed 30 seconds.
For melt analysis refer to instrument instructions.
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