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SF1ABIKB

Roche

KAPA SYBR® FAST One-Step

suitable for RT-PCR, 2 ×

NACRES:
NA.55

Quality Level

shelf life

≤6 mo.

packaging

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

concentration

2 ×

application(s)

RT-PCR: suitable

compatibility

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

shipped in

dry ice

storage temp.

−20°C

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General description

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.

Application

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

Biochem/physiol Actions

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°C. The RNase Inhibitor safeguards against degradation of RNA target due to RNase contamination.

Features and Benefits

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°C for 5 min.
  • 3 min at 95°C 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°C.
  • Do not exceed 25 μL reaction volumes.

Quality

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.

Preparation Note

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°C. The KAPA RT Mix is temperaturesensitive, and should be stored at -20°C 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°C when not in use. When protected from light, reagents are stable in the dark at 4°C 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.

Other Notes

For Research Use Only. Not for use in diagnostic procedures.

Legal Information

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

Kit Components Only

Product No.
Description

  • qPCR Master Mix (2X) with ROX High incorporated

  • dUTP

  • 50X KAPA RT Mix

Pictograms

Health hazard

Signal Word

Warning

Hazard Statements

hazcat

STOT SE 2

storage_class_code

12 - Non Combustible Liquids

WGK Germany

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

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