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KCQS01

Sigma-Aldrich

KiCqStart® SYBR® Green qPCR ReadyMix

Low ROX, for ABI and Stratagene 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 7500 Fast
for use with ABI 7500
for use with ABI ViiA 7
for use with Agilent AriaMx
for use with Douglas Scientific IntelliQube
for use with Qiagen Rotor-Gene Q
for use with QuantStudio
for use with Strategene Mx3000P
for use with Strategene Mx3005P
for use with Strategene Mx4000

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® SYBR® Green qPCR ReadyMix has been used:
  • for the amplification and quantification of DNA in real-time PCR (qPCR) assay
  • in the amplification and quantification of transcripts in 2-step quantitative reverse transcription polymerase chain reaction (qRT-PCR)
  • in the amplification of complementary DNA (cDNA) by real-time PCR (qPCR) assay
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

Quality

Kit components are free of contaminating DNase and RNase. KiCqStart® SYBR® Green qPCR ReadyMix, Low ROX is functionally tested in qPCR. Kinetic analysis must demonstrate linear resolution over six orders of dynamic range (r2 > 0.995) and a PCR efficiency > 90%.

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

6538 is a trademark of American Type Culture Collection
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.
Mx3000P is a registered trademark of Agilent Technologies, Inc.
Mx3005P is a registered trademark of Agilent Technologies, Inc.
Mx4000 is a trademark of Agilent Technologies, Inc.
QuantStudio is a trademark of IROA Technologies LLC
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
Rotor-Gene is a registered trademark of Qiagen GmbH
SYBR is a registered trademark of Life Technologies

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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Plasmodium falciparum EPCR-binding PfEMP1 expression increases with malaria disease severity and is elevated in retinopathy negative cerebral malaria
<BIG><BIG>Shabani E, et al.</BIG></BIG>
BMC Medicine, 15, 183-183 (2017)
Jemere Bekele Harito et al.
Water research, 114, 228-236 (2017-03-02)
Although standard methods for analyzing water samples for the protozoan parasites Cryptosporidium spp. and Giardia duodenalis are available and widely used, equivalent methods for analyzing water samples for Toxoplasma gondii oocysts are lacking. This is partly due to the lack
Estela Shabani et al.
BMC medicine, 15(1), 183-183 (2017-10-14)
Expression of group A and the A-like subset of group B Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is associated with severe malaria (SM). The diversity of var sequences combined with the challenges of distinct classification of patient pathologies has
Engineered zinc-finger transcription factors activate OCT4 (POU5F1), SOX2, KLF4, c-MYC (MYC) and miR302/367
<BIG><BIG>Ji Q, et al.</BIG></BIG>
Nucleic Acids Research, 42, 6158-6167 (2014)
Jemere Bekele Harito et al.
Water research, 127, 68-76 (2017-10-17)
Proof-of-principle of lectin-magnetic separation (LMS) for isolating Toxoplasma oocysts (pre-treated with 0.5% acidified pepsin (AP)) from water for subsequent detection by microscopy or molecular methods has been shown. However, application of this technique in the routine water-analysis laboratory requires that

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

Sigma’s Imprint RNA Immunoprecipitation Kit was used to copurify human argonaute 2 (Ago2)-associated RNAs from HeLa cells. MicroRNAs reverse transcribed and quantitating using Mysticq reagents.

Protocols

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

Analysis of gene expression data requires a stable reference or loading control. This reference is usually one or more reference genes.

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.

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.

See All

Related Content

SYBR® Green I, a commonly used fluorescent DNA binding dye, binds all double-stranded DNA and detection is monitored by measuring the increase in fluorescence throughout the cycle. Explore our LuminoCt® and KiCqStart® products for Fast qPCR or JumpStart™ reagents for conventional qPCR

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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