Taq PCR Master Mix Kit

For convenient PCR setup using a premixed solution

S_1281_3_LS_OEM_Taq_PCR_Master_Mix_Kit_1000U
Need bulk, customized or optimized products for commercial purposes? We also offer support with logistics, compliance and more. Reach out to cooperate with QIAGEN Strategic Partnerships & OEM

Taq PCR Master Mix Kit (1000 U)

Cat. No. / ID:   201445

12 x 1.7 ml Taq PCR Master Mix containing 1000 units Taq DNA Polymerase, 12 x 1.7 ml Distilled water
Quantity
1000 U
250 U
The Taq PCR Master Mix Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Need bulk, customized or optimized products for commercial purposes? We also offer support with logistics, compliance and more. Reach out to cooperate with QIAGEN Strategic Partnerships & OEM

Features

  • Minimal optimization due to QIAGEN PCR Buffer
  • Master mix format for easy reaction setup
  • Fewer pipetting steps minimize the risk of contamination

Product Details

Taq PCR Master Mix contains Taq DNA Polymerase, the unique QIAGEN PCR Buffer that minimizes the requirement for optimization, and dNTPs. The convenient 2x master mix format reduces pipetting steps, increasing throughput and reproducibility, while reducing the risk of contamination.

Performance

TheTaq PCR Master Mix Kit outperformed kits tested from other suppliers and ensures reliable PCR performance in a wide range of PCR applications — without the need for time-consuming optimization (see figure " Reproducible PCR"). Taq DNA Polymerase ensures highly specific amplification with different primer–template systems (see figures " Tolerance of different primer Tm values" and " Specific amplification of long PCR products"). Every lot of QIAGEN's Taq DNA Polymerase is subjected to a comprehensive range of quality control tests, including a stringent PCR specificity and reproducibility assay in which low-copy targets are amplified from human genomic DNA (see figure " Lot-to-lot reproducibility"). Due to the unique PCR buffer included in the master mix, optimization of PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced and often not required (see figures " Wide annealing temperature window" and " Tolerance to variable magnesium concentration").
Taq DNA Polymerase specifications

Concentration: 5 units/µl
Recombinant enzyme: Yes
Substrate analogs: dNTP, ddNTP, dUTP, biotin-11-dUTP, DIG-11-dUTP, fluorescent-dNTP/ddNTP
Extension rate: 2–4 kb/min at 72°C
Half-life: 10 min at 97°C; 60 min at 94°C
Amplification efficiency: ≥105 fold
5'–>3' exonuclease activity: Yes
Extra A addition: Yes
3'–>5' exonuclease activity: No
Contaminating nucleases: No
Contaminating RNases: No
Contaminating proteases: No
Self-priming activity: No

See figures

Principle

The Taq PCR Master Mix Kit includes QIAGEN's Taq DNA Polymerase in a premixed format. This ready-to-use solution also includes the QIAGEN PCR Buffer, MgCl2, and ultrapure dNTPs at optimized concentrations. Only primers and template DNA need to be added to set up PCR. Due to the convenient master mix format, pipetting errors are minimized, ensuring highly reproducible PCR results (see figure " Reproducible PCR"). Taq PCR Master Mix can be stored at 2–8°C for up to 2 months , allowing even faster PCR setup by eliminating thawing time.

Taq DNA Polymerase

TaqDNA Polymerase DNA Polymerase is a high-quality recombinant enzyme that is suitable for general and specialized PCR applications (see figures " Tolerance of different primer Tm values" and " Specific amplification of long PCR products").

QIAGEN PCR Buffer 

The innovative QIAGEN PCR Buffer has been developed to save time and effort by reducing the need for PCR optimization. QIAGEN PCR Buffer contains both KCl and (NH4)2SO4 (see figure " Increased specificity of primer annealing"). This unique buffer facilitates the amplification of specific PCR  products. During the annealing step of every PCR cycle, the buffer allows a high ratio of specific-to-nonspecific primer binding. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced and often not required (see figures " Wide annealing temperature window" and " Tolerance to variable magnesium concentration").

See figures

Procedure

Taq PCR Master Mix includesTaq DNA Polymerase, QIAGEN PCR Buffer, MgCl2, and ultrapure dNTPs at optimized concentrations. The ready-to-use 2x master mix format minimizes pipetting errors and also provides greater convenience. PCR setup is fast, easy, and straightforward — only primers and template DNA need to be added. The easy-to-follow protocol provided with the kit ensures specific amplification and PCR success at the first attempt.

Applications

The Taq PCR Master Mix Kit is used for standard and specialized applications, including:

  • General PCR
  • RT-PCR
  • Screening
  • PCR-based DNA fingerprinting (VNTR, STR, and RAPD)

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, RT-PCR, DNA fingerprinting
dNTP's includedYes (in Master Mix)
MastermixYes
Reaction typePCR amplification
Enzyme activity5' -> 3' exonuclease activity
Real-time or endpointEndpoint
Sample/target typeGenomic DNA and cDNA
Single or multiplexSingle
With/without hotstartWithout hotstart

Resources

Brochures & Guides (4)
Second edition — innovative tools
PCR 実験における重要項目と新技術
Addressing critical factors and new solutions
Safety Data Sheets (1)
Quick-Start Protocols (1)
Kit Handbooks (1)
For standard and specialized PCR applications with minimal optimization
Certificates of Analysis (1)

Publications

Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona.
Eremeeva ME; Bosserman EA; Demma LJ; Zambrano ML; Blau DM; Dasch GA;
Appl Environ Microbiol; 2006; 72 (8):5569-77 2006 Aug PMID:16885311
Population dynamics within a microbial consortium during growth on diesel fuel in saline environments.
Kleinsteuber S; Riis V; Fetzer I; Harms H; Müller S;
Appl Environ Microbiol; 2006; 72 (5):3531-42 2006 May PMID:16672500
PCR-based tandem epitope tagging system for Escherichia coli genome engineering.
Cho BK; Knight EM; Palsson BO;
Biotechniques; 2006; 40 (1):67-72 2006 Jan PMID:16454042
Genetic analysis of RpL38 and RpL5, two minute genes located in the centric heterochromatin of chromosome 2 of Drosophila melanogaster.
Marygold SJ; Coelho CM; Leevers SJ;
Genetics; 2004; 169 (2):683-95 2004 Nov 1 PMID:15520262
Cyclin C/cdk3 promotes Rb-dependent G0 exit.
Ren S; Rollins BJ;
Cell; 2004; 117 (2):239-51 2004 Apr 16 PMID:15084261

FAQ

What should the starting template DNA quality and quantity be for PCR?

Both the quality and quantity of nucleic acid starting template affect PCR, in particular the sensitivity and efficiency of amplification. PCR sensitivity and efficiency can be reduced by the presence of impurities in nucleic acid preparations or in biological samples. These PCR inhibitors are completely removed when template is prepared using QIAGEN Kits for nucleic acid purification. Please refer to the Brochure "Maximizing PCR and RT-PCR success" for additional information.

The optimal primer–template ratio has to be determined empirically. If too little template is used, primers may not be able to find their complementary sequences. Too much template may lead to an increase in mispriming events. Generally, no more than 1 ug of template DNA should be used per PCR reaction. As an initial guide, spectrophotometric and molar conversion values for different nucleic acid templates are listed below.

 

Spectrophotometric conversions for nucleic acid templates

1 A260 unit* Concentration (ug/ml)
Double-stranded DNA 50
Single-stranded DNA 33
Single-stranded RNA 40

*Absorbance at 260 nm = 1

 

Molar conversions for nucleic acid templates

Nucleic Acid Size pmol/ug Molecules/ug
1 kb DNA 1000 bp 1.52 9.1 x 1011
pUC 19 DNA 2686 bp 0.57 3.4 x 1011
pTZ18R DNA 2870 bp 0.54 3.2 x 1011
pBluescript II DNA 2961 bp 0.52 3.1 x 1011
Lambda DNA 48,502 bp 0.03 1.8 x 1010
Average mRNA 1930 nt 1.67 1.0 x 1012
Genomic DNA      
Escherichia coli 4.7 x 106* 3.0 x 10-4 1.8 x 108**
Drosophila melanogaster 1.4 x 108* 1.1 x 10-5 6.6 x 105**
Mus musculus (mouse) 2.7 x 109* 5.7 x 10-7 3.4 x 105**
Homo sapiens (human) 3.3 x 109* 4.7 x 10-7 2.8 x 105**

* Base pairs per haploid genome

** For single-copy genes

FAQ ID -74
What kind of PCR products can be cloned with the QIAGEN PCR Cloning Kit?

PCR products that will be cloned using the QIAGEN PCR Cloning Kit should be generated using a thermostable DNA Polymerase without proofreading activity, such as Taq DNA Polymerase. Such polymerases attach a single A overhang to their reaction products, which can hybridize to the U overhang of the pDrive Cloning Vector. For efficient addition of an A overhang during the PCR procedure, we recommend a final extension step for 10 min at 72°C as described in the standard protocols of the Taq PCR- and HotStarTaq PCR handbook.


 

FAQ ID -165
Do you have a protocol for polyacrylamide gel analysis of oligonucleotides?
Yes, please follow the Supplementary Protocol 'Polyacrylamide_gel_analysis_of_oligonucleotides' (PCR03).
FAQ ID -961
What makes QIAGEN's 10x Taq and HotStarTaq DNA Polymerase PCR buffer superior?
The QIAGEN 10x Taq and HotStarTaq DNA Polymerase PCR buffer contains a uniquely balanced combination of KCl and (NH4)2SO4. It provides stringent primer-annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers.
FAQ ID -566
How comparable is CoralLoad gel loading dye contained in various QIAGEN PCR Kits to Sigma Red?

CoralLoad gel tracking dye contained in Taq, HotStarTaq, TopTaq DNA Polymerase and TopTaq Master Mix Kits separates into 2 fragment-size dependent colors (orange and red) when loaded onto an agarose gel. Sigma Red buffer only has one color which is harder to visualize.

 

 

FAQ ID -1644
Have you tested the effect of inhibitors on PCR performance?

Yes. Please see Table 3 in our brochure Maximizing PCR and RT-PCR success. We tested the effects of different inhibitory substances in a number of PCR systems. We also analyzed the effect of including different volumes of reverse transcription (RT) reaction mixtures in PCR. Please see the table below for a list of commonly encountered template impurities and their inhibitory effects on PCR.

 

Impurities showing inhibitory effects on PCR

Substance Inhibitory concentration
SDS >0.005% (w/v)
Phenol >0.2% (v/v)
Ethanol >1% (v/v)
Isopropanol >1% (v/v)
Sodium Acetate ≥5 mM
Sodium Chloride ≥25 nM
EDTA ≥0.5 mM
Hemoglobin ≥1 mg/ml
Heparin ≥0.15 i.U./ml
Urea >20 mM
RT reaction mixture ≥15%

 

 

FAQ ID -818
Can QIAGEN's Taq- and HotstarTaq DNA Polymerases be used for cycle sequencing?
Taq DNA Polymerase and HotStarTaq DNA Polymerase are compatible with cycle sequencing. However, our buffer system is not optimized for this purpose. Optimization of reaction conditions is therefore required when using these Polymerases for cycle sequencing. Unfortunately, we do not have any protocols for this application. An initial activation of the enzyme is necessary if HotStarTaq DNA Polymerase is used.
FAQ ID -741
How can one determine the optimal annealing temperature for a specific PCR assay?

To determine the optimal annealing temperature for a PCR assay, a Temperature Gradient experiment should be performed. To do this, you will set up several PCR reactions in duplicate for the same primer/template combination, using the same PCR chemistry, and subject each of the reactions to a slightly different annealing temperature within a specified range. If a thermal cycler with a temperature gradient function can be used, you can simply program a temperature range for adjacent wells in the cycling block. If no cycler with a gradient function exists in your lab, you will either have to perform duplicate reactions at different temperatures in different machines (if available), or back to back in the same machine.

 

FAQ ID -288