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KCQS02

Sigma-Aldrich

KiCqStart® SYBR® Green qPCR ReadyMix

with ROX for ABI instruments

Synonym(s):

qPCR master mix, sybr green qPCR

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About This Item

UNSPSC Code:
41106300
NACRES:
NA.55

form

liquid

usage

sufficient for 1250 reactions
sufficient for 250 reactions
sufficient for 5000 reactions

feature

dNTPs included
hotstart

storage condition

protect from light

technique(s)

qPCR: suitable

color

colorless

input

purified DNA

compatibility

for use with ABI 5700
for use with ABI 7000
for use with ABI 7300
for use with ABI 7700
for use with ABI 7900 HT Fast
for use with ABI 7900 HT
for use with ABI 7900
for use with ABI StepOne
for use with ABI StepOnePlus

detection method

SYBR® Green

shipped in

dry ice

storage temp.

−20°C

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General description

KiCqStart SYBR Green qPCR ReadyMix is a 2X concentrated, ready-to-use master mix that contains all components, except primers and template, for real-time quantitative PCR (qPCR) This unique combination of proprietary buffer, stabilizers, and Hot-Start Taq DNA polymerase delivers maximum PCR efficiency, sensitivity, specificity and robust fluorescent signal using fast, or conventional, cycling protocols with SYBR Green qPCR.

Highly specific amplification is crucial to successful qPCR with SYBR Green I dye technology because this dye binds to and detects any dsDNA generated during amplification. Hot-Start Taq DNA polymerase is antibody mediated to be inactive prior to the initial PCR denaturation step.

Application

KiCqStart® SYBRSYB® Green qPCR ReadyMix has been used:

  • in the amplification and quantification of cDNA reverse transcribed from RNA extracted from mice brain samples in a 2-step RT-qPCR assay
  • to analyze DNA purified by ChIP technique
  • to perform gene expression analysis
  • in amplification of RNA isolated from hearts of adult TL wild-type fish by quantitative real-time polymerase chain reaction (PCR)
PCR applications:
  • Gene expression
  • DNA quantification
  • CHiP

Features and Benefits

  • Assay results in as little as 33 minutes
  • Highly efficient and sensitive real-time PCR results
  • Little/no optimization required

Components

2X reaction buffer containing optimized concentrations of MgCl2, dNTPs, (dATP, dCTP, dGTP, dTTP), KiCqStart Taq DNA Polymerase, SYBR Green dye, ROX Reference Dye (for 580-585 nm excitation), and stabilizers.

packaging:
250 reactions* = 2 X 1.25 mL tubes
1250 reactions* = 10 X 1.25 mL tubes
5000 reactions* = 1 X 50 mL tube
*number of reactions based on a 20uL volume

Other Notes

Storage Conditions:
KiCqStart SYBR Green qPCR ReadyMix is stable for 1 year when stored in a constant temperature freezer at -20°C, protected from light. For convenience, it may be stored unfrozen at +2 to +8°C for up to 6 months. After thawing, mix thoroughly before using. Repeated freezing and thawing of the product is not recommended. However, the product demonstrated no loss of performance after 20 freeze-thaw cycles or 2 months at +20°C.

Legal Information

Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
KiCqStart is a registered trademark of QIAGEN Beverly Inc.
ROX is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
ReadyMix is a trademark of Sigma-Aldrich Co. 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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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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Gene expression study of phase I and II metabolizing enzymes in RPTEC/TERT1 cell line: application in in vitro nephrotoxicity prediction.
Shah H
Xenobiotica, 3, 1-7 (2016)
Gene expression study of phase I and II metabolizing enzymes in RPTEC/TERT1 cell line: application in in vitro nephrotoxicity prediction
<BIG><BIG>Shah H, et al.</BIG></BIG>
Xenobiotica, 47, 837-843 (2017)
Aneesh K Ramaswamy et al.
Matrix biology plus, 4, 100014-100014 (2019-09-04)
Elastogenesis within the medial layer of the aortic wall involves a cascade of events orchestrated primarily by smooth muscle cells, including transcription of elastin and a cadre of elastin chaperone matricellular proteins, deposition and cross-linking of tropoelastin coacervates, and maturation
Eoghan M Cunnane et al.
Bioengineering (Basel, Switzerland), 8(5) (2021-05-01)
Macromolecular components of the vascular extracellular matrix (ECM), particularly elastic fibers and collagen fibers, are critical for the proper physiological function of arteries. When the unique biomechanical combination of these fibers is disrupted, or in the ultimate extreme where fibers
The functional and inflammatory response of brain endothelial cells to toll-like receptor agonists
Johnson RH, et al.
Scientific Reports, 8, 1-12 (2018)

Articles

After a traditional PCR has been completed, the PCR/qPCR data analysis is conducted by resolution through an agarose gel or, more recently, through a capillary.

PCR assay guide navigates you through primer validation and other assay optimization factors to ensure high sensitivity and specificity for optimum DNA/ RNA quantification.

Real-time polymerase chain reaction allows researchers to estimate the quantity of starting material in a sample. It has a much wider dynamic range of analysis than conventional PCR

Protocols

Quantitative PCR protocol using SYBR Green reagents. Procedure supports most qPCR instruments.

Gradient PCR for assay optimization is to determine the optimum annealing temperature (Ta) of the primers by testing identical reactions containing a fixed primer concentration, across a range of annealing temperatures.

Measuring a target quantity relative to one or more stable reference genes using SYBR Green I dye detection is a common application of qPCR. Below is a standard protocol that can be adapted to specific experimental needs.

Once an assay has been optimized, it is important to verify the reaction efficiency. This information is important when reporting and comparing assays. In this example protocol, the assay efficiency is compared over a wide and narrow dynamic range of cDNA concentrations.

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