Taq PCR Core Kit

用于常规和特殊的PCR，包含dNTP混合液

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Taq PCR Core Kit (1000 U)

Cat. No. / ID: 201225

4 x 250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl₂, dNTP Mix

Quantity

1000 U

250 U

Inquire

Taq PCR Core 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
独特的即用型PCR缓冲液，操作更快速简单
Q-Solution可帮助扩增GC含量高的模板
提供多种规格的包装方便使用

产品详情

Taq PCR Core Kit包含Taq DNA Polymerase、独特的QIAGEN PCR Buffer能最大限度地减少使用时对PCR条件的优化，以及dNTP混合液。同时还提供新型的辅助剂Q-Solution，有助于实现某些“困难”模板（比如，高GC含量的模板）的高效扩增。CoralLoad PCR Buffer（包括两种胶示踪染料）使PCR产物可以直接上样电泳。

绩效

Taq PCR Core Kit在大量的PCR应用中有优秀的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和特殊的PCR应用（参见" Tolerance of different primer T_m Values"和" Specific amplification of long PCR products"）。每批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" Lot-to-lot reproducibility"）。试剂盒中还包含独特制备的QIAGEN PCR Buffer和CoralLoad PCR Buffer，能在最少的优化过程的PCR条件下，进行高特异性的PCR（参见" Wide annealing-temperature window"和" Tolerance to variable magnesium concentration"）。此外，CoralLoad PCR Buffer能使PCR产物直接上样到琼脂糖凝胶，操作简单，能更快得到结果。这次的PCR可通过使用试剂盒内的PCR辅助剂Q-Solution进行优化（参见" Amplification of difficult templates"）。

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

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.

Amplification of difficult templates.

原理

Taq PCR Core Kit包含所有简单可靠的PCR所需的所有试剂，Taq DNA Polymerase、QIAGEN PCR Buffer、CoralLoad PCR Buffer、Q-Solution、dNTP Mix和MgCl₂.

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独特的均衡结合，相比传统的PCR缓冲液，该PCR缓冲液提供大范围退火温度和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"）。

CoralLoad PCR Buffer

CoralLoad PCR Buffer具有所有QIAGEN PCR Buffer的优点。此外，还可以直接将PCR反应物上样到琼脂糖凝胶上，无需额外单独的凝胶上样缓冲液。CoralLoad PCR Buffer与传统的QIAGEN PCR Buffer一样能提供高PCR特异性和最小的反应优化条件。而且该缓冲液还包含两种标记染料，橙色染料和红色染料，有助于消除DNA迁移距离并优化琼糖脂凝胶跑样时间（参见" CoralLoad PCR Buffer"）。该缓冲液优化了移液的可视性，并能够直接将PCR产物上样到凝胶上，便于操作。

Q-Solution

Q-Solution有利于通过修改DNA熔化行为扩增高GC含量的模板或具有高度二级结构的模板。使用该特殊的试剂通常能够进行或提高欠佳的PCR（参见" Amplification of difficult templates"）。不同于DMSO和其他的PCR辅助剂，Q-Solution用于各种引物模板的特定浓度，且无毒。

查看图表

Tolerance of different primer Tm values.

Specific amplification of long PCR products.

Increased specificity of primer annealing.

Tolerance to variable magnesium concentration.

CoralLoad PCR Buffer.

Amplification of difficult templates.

程序

Taq PCR Core Kit提供所有构建PCR反应体系和用于多重基于PCR应用的所有组件。该试剂盒提供经优化、易于使用、精简的实验方案，确保获得成功的PCR结果。该试剂盒中还包含一款独特的PCR辅助剂，Q-Solution，用于简化欠佳的PCR反应。

应用

Taq DNA Polymerase用于常规的和特殊的应用，包括：

常规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

CoralLoad PCR Buffer.

Specifications

Features	Specifications
ApplicationsZH	PCR, RT-PCR, DNA fingerprinting
Sample/target type	Genomic DNA and cDNA
Real-time or endpoint	Endpoint
dNTP's included	Yes
Single or multiplex	Single
Reaction type	PCR amplification
Mastermix	No
With/without hotstart	Without hotstart
Enzyme activity	5' -> 3' exonuclease activity

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

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

No, we do not sell Q-Solution separately. It is available only as a component of the Taq DNA Polymerase, Taq PCR Core, HotStarTaq DNA Polymerase, QIAGEN Multiplex PCR-, and the QIAGEN OneStep RT-PCR Kits.

FAQ ID -204

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

FAQ ID -961

Taq DNA Polymerase has an intrinsic RNA-dependent DNA polymerase activity (reverse transcriptase activity). However, this activity is very low and is only present under buffer conditions that are completely different from those present during PCR. Therefore, "no-RT" controls would not give false positive results due to reverse transcription activity of the Taq polymerase.

FAQ ID -523

Not necessarily. In a lot of cases, the uniquely formulated PCR Buffer provided in the HotStarTag Plus DNA Polymerase, HotStar HiFidelity Polymerase, Taq DNA Polymerase, HotStarTaq DNA Polymerase, and QIAGEN Multiplex PCR Kits provides optimal amplification of specific PCR products. The usefulness of Q-Solution needs to be determined empirically for each primer/template setup, by running parallel PCR reactions with and without Q-Solution under the same cycling conditions.

Q-Solution changes the melting behavior of DNA and will often improve a suboptimal PCR caused by templates that have a high degree of secondary structure or high GC-contents. For more details on the effects of Q-Solution on PCR amplification, please see the Q-Solution sections of the HotStarTag Plus DNA Polymerase, HotStar HiFidelity Polymerase, Taq DNA Polymerase, HotStarTaq DNA Polymerase, and the QIAGEN Multiplex PCR Handbooks.

FAQ ID -380

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

QIAGEN's 10x PCR Buffer provided in the Taq DNA Polymerase, Taq PCR Core, and HotStarTaq DNA Polymerase Kits contains:

Tris-Cl
KCl
(NH₄)₂SO₄
15 mM MgCl₂ ; at pH 8.7 (20°C).

Note that further details on the composition of the 10x PCR Buffer are proprietary.

FAQ ID -606

The DNA yield obtained in a PCR reaction depends on the size of the amplicon, design of the primers, starting amount of template and primers, amplification efficiency, reaction volume, numbers of PCR cycles etc. Therefore it is really difficult to predict what yield to expect. Nevertheless, in our experience, approximately 1 µg is a good guess for most cases.

FAQ ID -750

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