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KAPA SYBR® FAST One-Step

suitable for RT-PCR, 2 ×, Universal

Synonym(s):

SYBR, qPCR

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

Quality Level

100

usage

sufficient for 100 reactions
 sufficient for 1000 reactions
 sufficient for 500 reactions

shelf life

≤6 mo.

feature

dNTPs included: no
hotstart: no

packaging

kit of 1 mL (100 x 20 μL rxn; KK4650)
kit of 10 mL (1000 x 20 μL rxn; KK4652)
kit of 5 mL (500 x 20 μL rxn; KK4651)

manufacturer/tradename

Roche

concentration

2 ×

technique(s)

RT-PCR: suitable
qPCR: suitable

input

purified DNA

compatibility

for use with Agilent Mx3000P
for use with Agilent Mx3005P
for use with Agilent Mx4000
for use with Applied Biosystems® 5700
for use with Applied Biosystems® 7000
for use with Applied Biosystems® 7300
for use with Applied Biosystems® 7500
for use with Applied Biosystems® 7700
for use with Applied Biosystems® 7900HT
for use with Applied Biosystems® ViiA 7
for use with QuantStudio™
for use with StepOne™
for use with StepOnePlus™

detection method

SYBR® Green

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
  • Gene expression analysis
  • Low-copy gene detection
  • Microarray validationmiRNA research
  • Gene knockdown validation
  • Real-time quantitative reverse transcription-PCR (qRT-PCR)
  • RNAi and miRNA research

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

Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
QuantStudio is a trademark of IROA Technologies LLC
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
  • KAPA RT Mix
  • dUTP
  • KAPA SYBR® FAST DNA Polymerase
  • reaction buffer
  • dNTPs
  • SYBR® Green I dye
  • MgCl2 2.5 mM

Pictograms

Health hazard
GHS08

Signal Word

Warning

Hazard Statements

H371

Hazard Classifications

STOT SE 2

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Yuxi Zhang et al.
Cell chemical biology, 24(4), 471-480 (2017-04-04)
Stimulating autophagy is a promising therapeutic strategy for slowing the progression of neurodegenerative disease. Neurons are insensitive to current approaches based on mTOR inhibition for activating autophagy, and instead may rely on the Parkinson's disease-associated proteins PINK1 and PARKIN to activate
Zharko Daniloski et al.
Cell, 184(1), 92-105 (2020-11-05)
To better understand host-virus genetic dependencies and find potential therapeutic targets for COVID-19, we performed a genome-scale CRISPR loss-of-function screen to identify host factors required for SARS-CoV-2 viral infection of human alveolar epithelial cells. Top-ranked genes cluster into distinct pathways
Federica Fontana et al.
Cell reports, 32(2), 107883-107883 (2020-07-16)
The formation of cardiac valves depends on mechanical forces exerted by blood flow. Endocardial cells lining the interior of the heart are sensitive to these stimuli and respond by rearranging into luminal cells subjected to shear stress and abluminal cells

Articles

Directed Evolution

An overview of directed evolution and the methods for generating proteins with optimized or entirely new functions.

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