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SF1BRKB

Roche

KAPA SYBR® FAST One-Step

Bio-Rad iCycler®

NACRES:
NA.55

Quality Level

shelf life

≤6 mo.

packaging

kit of 1 mL (100 x 20 μL rxn; KK4670)
kit of 10 mL (1000 x 20 μL rxn; KK4672)
kit of 5 mL (500 x 20 μL rxn; KK4671)

manufacturer/tradename

Roche

compatibility

for use with Bio-Rad iCycler

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® qRT-PCR Master Mix (2X) Bio-Rad® iCycler® is suitable for:
  • Real-time polymerase chain reaction (RT-PCR)
  • Gene expression analysis
  • Low-copy gene detection
  • Microarray validation
  • miRNA research
  • Gene knockdown validation

Biochem/physiol Actions

KAPA SYBR® FAST DNA Polymerase is inactive 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. The RNase Inhibitor averts degradation of RNA target due to RNase contamination. Moloney murine leukemia virus reverse transcriptase (M-MuLV RT) has a high affinity for RNA and is optimized for cDNA synthesis at 42°C.

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

Bio-Rad is a registered trademark of Bio-Rad Laboratories, Inc.
OneRack is a registered trademark of Heathrow Scientific LLC
SYBR is a registered trademark of Life Technologies
iCycler is a registered trademark of Bio-Rad

Kit Components Only

Product No.
Description

  • qPCR Master Mix (2X) with fluorescein incorporated

  • dUTP 10 mM

  • 50X KAPA RT Mix

Certificate of Analysis

Certificate of Origin

A novel CBF that regulates abiotic stress response and the ripening process in oil palm (Elaeis?guineensis) fruits
Ebrahimi M, et al.
Tree genetics & genomes, 11(3), 56-56 (2015)
Articles
An overview of directed evolution and the methods for generating proteins with optimized or entirely new functions.
Read More

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