产品说明
一般描述
KAPA SYBR FAST One-Step qRT-PCR Kits are designed for optimal performance in stringent qRT-PCR reaction conditions, exhibiting dramatic improvements in sensitivity, specificity, reaction efficiency, and fluorescence. The kit is optimized for rapid one-step, one-tube RNA quantification, reducing experimental variation and contamination with a convenient qRT-PCR protocol.
KAPA SYBR FAST One-Step qRT-PCR Kits contain M-MuLV reverse transcriptase, RNase inhibitor, and a novel DNA polymerase evolved via directed evolution.
应用
KAPA SYBR® FAST One-Step is suitable for:
- One-step quantitative reverse transcriptase PCR (qRT-PCR)
- Gene expression analysis
- Low-copy gene detection
- Microarray validation
- miRNA research
- Gene knockdown validation
生化/生理作用
KAPA SYBR® FAST DNA Polymerase displays no enzymatic activity at ambient temperature. This prevents the formation of misprimed products and primer-dimers during reaction setup prior to the first denaturation step and results in high PCR specificity and accurate quantification. Moloney murine leukemia virus reverse transcriptase (M-MuLV-RT) has a high affinity for RNA and is optimized for cDNA synthesis at 42℃. The RNase Inhibitor safeguards against degradation of RNA target due to RNase contamination.
特点和优势
Increase signal and reaction efficiency:
- Consistently higher amplification efficiencies for accurate qPCR
- Higher fluorescence, earlier Cq values and improved reaction efficiencies
Amplify across a broad range of target lengths:
- Robust performance independent of amplicon size
Improve sensitivity and reproducibility:
- Accurate interrogation across a wide range of RNA template concentrations
Quick Notes:
- This kit contains wild-type M-MuLV and an engineered enzyme optimized for qPCR using SYBR Green I dye chemistry.
- The 2X master mix contains a proprietary buffer. Together with the novel enzyme, this improves amplification efficiency of both GC- and AT-rich targets.
- Use only gene-specific primers for one-step qRT-PCR.
- Optimal cDNA synthesis is achieved at 42℃ for 5 min.
- 3 min at 95℃ is sufficient for RT inactivation and DNA polymerase activation.
- For 3-step cycling, use 20 sec for primer annealing and 1 sec for extension/data acquisition at 72℃.
- Do not exceed 25 μL reaction volumes.
质量
All kit components are subjected to stringent functional quality control, are free of detectable contaminating exo and endonuclease activity, and meet strict requirements with respect to DNA contamination.
制备说明
Always ensure that components have been fully thawed and thoroughly mixed before use. The KAPA SYBR FAST qPCR Master Mix (2X) may not freeze solidly, even when stored at -20℃. The KAPA RT Mix is temperaturesensitive, and should be stored at -20℃ and kept on ice during use.
The SYBR Green I dye contained in the KAPA SYBR FAST qPCR Master Mix (2X) and ROX/fluorescein dyes (depending on kit configuration) are light sensitive. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity.
KAPA SYBR FAST qPCR Master Mix (2X) is stable through 30 freeze-thaw cycles. Ensure that all reagents are stored protected from light at -20℃ when not in use. When protected from light, reagents are stable in the dark at 4℃ for at least one week and may be stored at this temperature for short-term use provided that they do not become contaminated with microbes and/or nucleases.
其他说明
For Research Use Only. Not for use in diagnostic procedures.
法律信息
ABI PRISM is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
SYBR is a registered trademark of Life Technologies
StepOne is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
StepOnePlus is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
基本信息
| NACRES | NA.55 |
产品性质
| 质量水平 | 100 |
| 用途 | sufficient for 100 reactions sufficient for 1000 reactions sufficient for 500 reactions |
| 保质期 | ≤6 mo. |
| 特点 | dNTPs included: no hotstart: no |
| 包装 | kit of 1 mL (100 x 20 μL rxn; KK4660) kit of 10 mL (1000 x 20 μL rxn; KK4662) kit of 5 mL (500 x 20 μL rxn; KK4661) pkg of |
| manufacturer/tradename | Roche |
| 浓度 | 2 × |
| technique(s) | RT-PCR: suitable qPCR: suitable |
| input | purified DNA |
| 相容性 | for use with Applied Biosystems® 5700 for use with Applied Biosystems® 7000 for use with Applied Biosystems® 7300 for use with Applied Biosystems® 7700 for use with Applied Biosystems® 7900HT for use with StepOne™ for use with StepOnePlus™ |
| detection method | SYBR® Green |
| 运输 | dry ice |
| 储存温度 | −20℃ |
组份列表
组份不可单独销售
| 货号 | 组份 |
| qPCR Master Mix (2X) with ROX™ High incorporated | |
| dUTP | |
| 50X KAPA RT Mix |
安全信息
| 象形图 |  |
| 警示用语： | Warning |
| 危险声明 | H371 |
| 预防措施声明 | P260 - P264 - P270 - P308 + P311 - P405 - P501 |
| 危险分类 | STOT SE 2 |
| 储存分类代码 | 12 - Non Combustible Liquids |
| WGK | WGK 1 |
| 闪点(F) | does not flash |
| 闪点(C) | does not flash |
