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Roche

KAPA SYBR® FAST

suitable for qPCR, 2 ×, for LightCycler®480

Synonym(s):

Lightcycler, qPCR

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

Quality Level

usage

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

shelf life

≤12 mo.

feature

dNTPs included
hotstart

packaging

kit of 1 mL (100 x 20 μL rxn; KK4609)
kit of 10 mL (1000 x 20 μL rxn; KK4611)
kit of 5 mL (500 x 20 μL rxn; KK4610)

manufacturer/tradename

Roche

concentration

2 ×

technique(s)

qPCR: suitable

input

purified DNA

compatibility

for use with Bio-Rad CFX96
for use with Bio-Rad Chromo4 (Real-Time Detector)
for use with Bio-Rad DNA Engine Opticon
for use with Bio-Rad Opticon 2
for use with Eppendorf® Mastercycler (ep realplex )
for use with Illumina Eco
for use with Roche LightCycler 1.5/2.0
for use with Roche LightCycler 480
for use with Roche LightCycler 96
for use with Roche LightCycler Nano
for use with Rotor-Gene®
for use with Smart Cycler®

detection method

SYBR® Green

storage temp.

−20°C

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General description

KAPA SYBR® FAST qPCR Kits contain the first DNA polymerase engineered via directed evolution to be more tolerant of SYBR Green I dye inhibition.

The improved robustness, processivity, and speed of KAPA SYBR FAST qPCR Kits result in consistently high amplification efficiencies enabling more accurate relative quantification for gene expression analysis. KAPA SYBR FAST qPCR Kits, developed to perform optimally in stringent real-time PCR reaction conditions, exhibit dramatic improvements to signal-to-noise ratio (fluorescence), quantification cycle (Cq), linearity, and sensitivity.

Application

KAPA SYBR® FAST for LightCycler®480 has been used for:
  • Gene expression
  • Gene knockdown validation
  • Microarray validation
  • ChIP analysis
  • Low copy detection
  • Absolute quantification of NGS libraries
  • Quantitative real-time PCR (qPCR)

Features and Benefits

Quantitate changes in gene expression more accurately
  • High reaction efficiency between 95 - 105% improves accuracy and reproducibility.
  • Unbiased efficiency across a wide range of GC contents and amplicon lengths.

Detect low copy and difficult targets consistently
  • Improved processivity results in earlier Cq scores.
  • Higher fluorescence detection across varying AT- and GC-rich targets.
  • Novel enzyme is resistant to SYBR® inhibition.

Complete real-time PCR runs in just 40 minutes
  • 55% shorter run times with fast cycling protocol.
  • Maintain high performance when switching from standard to fast protocols.

Quick Notes:
  • This kit contains 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.
  • 20 sec initial denaturation at 95°C is sufficient for enzyme activation. When working with complex templates, an initial denaturation of 3 min is recommended.
  • 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. KAPA SYBR® FAST qPCR Master Mix (2X) may not freeze solidly, even when stored at -20°C.

The SYBR Green I dye contained in 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

Eppendorf is a registered trademark of Eppendorf AG
LightCycler is a registered trademark of Roche
Rotor-Gene is a registered trademark of Qiagen GmbH
SYBR is a registered trademark of Life Technologies
SmartCycler is a registered trademark of Cepheid, Inc.

Kit Components Only

Product No.
Description

  • KAPA SYBR® FAST DNA Polymerase

  • reaction buffer

  • dNTPs

  • SYBR® Green I dye

  • MgCl2 2.5 mM

Pictograms

Health hazard

Signal Word

Warning

Hazard Statements

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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Articles

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

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