Taq PCR Master Mix Kit

편리한 PCR setup을 위한 premixed solution

상업적인 목적을 위해 제품이 대량으로 필요하거나 맞춤형 또는 최적화 제품이 필요하십니까? QIAGEN에서는 물류, 규정 준수 등에 대한 지원도 제공합니다. QIAGEN에 문의하여 전략적 파트너십과 OEM을 통해 협력하는 방법을 알아보세요

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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

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Taq PCR Master Mix Kit은/는 분자생물학 분야에 사용하기 위한 것입니다. 이 제품은 질병의 진단, 예방, 또는 치료용이 아닙니다.

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특징

QIAGEN PCR Buffer for minimal optimization
쉽고 간편한 조작을위해 format 선택이 가능함
Fewer pipetting steps minimize the risk of contamination

제품 세부 정보

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.

성능

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 ""). Taq DNA Polymerase ensures highly specific amplification with different primer–template systems (see figures "" and ""). 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 ""). 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 "" and "").

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

그림 참조

Reproducible PCR.

Tolerance of different primer Tm values.

Specific amplification of long PCR products.

Lot-to-lot reproducibility.

Tolerance to variable magnesium concentration.

원리

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

Taq DNA Polymerase DNA Polymerase is a high-quality recombinant enzyme that is suitable for general and specialized PCR applications (see figures " Tolerance of different primer T_m 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").

그림 참조

Reproducible PCR.

Tolerance of different primer Tm values.

Specific amplification of long PCR products.

Increased specificity of primer annealing.

Tolerance to variable magnesium concentration.

절차

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.

응용 분야

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)

지원되는 데이터 및 수치

Reproducible PCR.

A fragment of the hepatitis B surface antigen gene (gene S) was amplified from 10, 20, and 50 copies of target template, using the Taq PCR Master Mix Kit. Five parallel amplifications were performed for each amount of starting template DNA. Equal volumes of the PCR products were analyzed on a 2% agarose gel. C: negative control; M: markers.

Specifications

Features	Specifications
Applications	PCR, RT-PCR, DNA fingerprinting
dNTP's included	Yes (in Master Mix)
Mastermix	Yes
Reaction type	PCR amplification
Enzyme activity	5' -> 3' exonuclease activity
Real-time or endpoint	Endpoint
Sample/target type	Genomic DNA and cDNA
Single or multiplex	Single
With/without hotstart	Without hotstart

리소스

Second edition — innovative tools

Addressing critical factors and new solutions

For standard and specialized PCR applications with minimal optimization

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

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

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

Yes, please follow the Supplementary Protocol 'Polyacrylamide_gel_analysis_of_oligonucleotides' (PCR03).

FAQ ID -961

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

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

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

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

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