Taq PCR Master Mix Kit

预混合的PCR反应液方便PCR体系构建

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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

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Quantity

1000 U

250 U

Taq PCR Master Mix Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

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特点

QIAGEN PCR缓冲体系最大限度地减少了PCR条件的优化
预混液规格易于反应体系构建
更少的移液操作最大限度减少污染风险

产品详情

Taq PCR Master Mix包含Taq DNA Polymerase、独特的QIAGEN PCR Buffer最大限度的减少优化需求，以及dNTP。方便的2倍预混液减少了移液步骤，提高了反应的通量和可重复性，同时降低了污染的风险。

绩效

Taq PCR Master Mix Kit相比其他供应商测试的试剂盒有更卓越的表现，并在大量的PCR应用中有可靠的PCR性能，无需耗时的优化过程。该试剂盒包含Taq DNA Polymerase，一种高品质的重组酶适用于常规的和特殊的PCR应用（参见""）。Taq DNA Polymerase确保在不同的引物模板体系中高特异性扩增（参见""和""）。每批QIAGEN's Taq DNA Polymerase受到全面地质量监控测试，包括严格的PCR特异性和重复性测试，在此测试中可从人类基因组DNA中扩增低拷贝靶（参见""）。由于预混液中还包含独特的PCR缓冲液，可通过改变退火温度或Mg²⁺浓度，即可显著降低优化过程，而且通常不需要优化（参见""和""）。

Taq DNA Polymerase规格

浓度：5单位/µl
重组酶：有
底物类似物：dNTP、ddNTP、dUTP、biotin-11-dUTP、DIG-11-dUTP、荧光-dNTP/ddNTP
延伸率：72°C下2–4 kb/分钟
半衰期：97°C下10分钟；94°C下60分钟
扩增效率：≥10⁵倍
5'–>3'外切酶活性：有
Extra A辅助剂: 有
3'–>5'外切酶活性：无
污染核酸：无
污染RNA酶：无
污染蛋白酶：无
自吸泵活性：无

查看图表

Reproducible PCR.

Tolerance of different primer Tm values.

Specific amplification of long PCR products.

Lot-to-lot reproducibility.

Increased specificity of primer annealing.

Tolerance to variable magnesium concentration.

原理

Taq PCR Master Mix Kit包含预混合规格的QIAGEN's Taq DNA Polymerase。这种易于使用的反应液同样包含QIAGEN PCR Buffer、MgCl₂和具优化浓度的超纯dNTP。只需加入引物和模板DNA即可构建PCR反应体系。由于方便的预混液规格，最大限度地减少了移液失误，确保了高重复的PCR结果（参见" Reproducible PCR"）。Taq PCR Master Mix可在2–8°C下储存长达2个月，通过去除解冻时间来构建更快速的PCR反应体系。

Taq DNA Polymerase

Taq DNA Polymerase是高品质的重组酶，适用于常规的和特殊的PCR应用（参见" Tolerance of different primer T_m values"和" Specific amplification of long PCR products"）。

QIAGEN PCR Buffer

研发新型的QIAGEN PCR Buffer通过减少使用时对PCR条件的优化，来节省时间和精力。QIAGEN PCR Buffer包括KCl和(NH₄)₂SO₄。独特的缓冲体系有助于特异性PCR产物的扩增。在每个PCR循环的退火步骤中，该缓冲液具有特异性-非特异性引物的高比率结合。由于KCl和(NH₄)₂SO₄独特的平衡结合，相比传统的PCR缓冲液，该PCR缓冲液提供更大范围退火温度和Mg²⁺浓度条件，以及严格的引物退火条件。通过改变退火温度或Mg²⁺浓度，即可显著减少PCR优化过程，而且通常不需要优化（参见" Wide annealing temperature window"和" Tolerance to variable magnesium concentration"）。

查看图表

Reproducible PCR.

Tolerance of different primer Tm values.

Specific amplification of long PCR products.

Increased specificity of primer annealing.

Tolerance to variable magnesium concentration.

程序

Taq PCR Master Mix包含Taq DNA Polymerase、QIAGEN PCR Buffer、MgCl₂和优化浓度的dNTP。即用型2倍预混液规格最大限度减少了移液食物，同时提供了极大便利。构建PCR反应体系快速、简单、直接，只需加入引物和模板DNA。该试剂盒还易于操作的实验方案，确保了在第一次操作中就可获得特异性扩增并成功进行PCR反应。

应用

Taq PCR Master Mix Kit适用于常规的和特殊的应用，包括：

常规的PCR
RT-PCR
筛选
基于PCR的DNA指纹图谱分析（VNTR、STR和RAPD）

辅助数据和图表

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

资源

Second edition — innovative tools

Addressing critical factors and new solutions

Download Safety Data Sheets for QIAGEN product components.

For standard and specialized PCR applications with minimal optimization

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