Tap DNA Polymerase

Product Information

Taq DNA Polymerase, abbreviated as Taq enzyme, catalyzes the polymerization of deoxyribonucleotides in a 5' to 3' direction dependent on a DNA template. Taq enzyme lacks 3' to 5' exonuclease activity and has very low 5' to 3' exonuclease activity. Due to this lack of proofreading capability, it functions similarly to a nucleotidyl transferase during DNA synthesis. The PCR products generated have a single adenine (A) overhang at the 3’ end, allowing direct cloning into T-vectors.

Applications

PCR, DNA labeling, and sequencing. It can amplify DNA fragments up to 5 kb in length, with the maximum amplifiable size reaching up to 8 kb. Typically suitable for fragments below 3 kb.
Nucleic acid labeling reactions. PCR fragments with A-overhangs at the 3’ end are suitable for cloning into T-vectors. Biotin, digoxigenin, or fluorescent-labeled dNTPs can be incorporated during PCR.
DNA sequencing.

Storage

Store at -20°C

Product Components

Component No.	Component Name	Specifications
HX1001-1	Taq DNA Polymerase (5 U/μl)	200 U	1000 U	5000 U
HX1001-2	10X PCR Buffer (with Mg²⁺)	1 ml	1 ml×4	10 ml×2

Inactivation or Inhibition: Phenol-chloroform extraction can inactivate Taq enzyme. Adding sodium deoxycholate to 0.06%, SDS to 0.01%, or Sarkosyl to 0.02% can also inhibit Taq enzyme activity.

Notes:

Avoid contamination from trace amounts of DNA intended for amplification. Consider including a no-template control to verify potential contamination.
The error rate of Taq DNA polymerase is approximately 2.2×10^-5 per cycle during PCR. For cloning DNA fragments larger than 1 kb, a high-fidelity DNA polymerase with a lower error rate is recommended.
Taq DNA polymerase is the best choice for standard PCR or RT-PCR qualitative and quantitative detection. For your safety and health, wear lab coat and disposable gloves when handling this product.

1. Thaw and mix all solutions required for the PCR reaction. Setting up the PCR reaction system:

Reagent	Final Concentration	Volume
Double-distilled water or Milli-Q water	-	(36.75-x) μl
10X PCR Buffer (with Mg2+)	1 X	5 μl
dNTP (2.5 mM each)	0.2 mM each	4 μl
Template DNA	10 pg - 1 μg	X μl
Primer Mix (10 μM each)	0.8 μM	4 μl
Taq DNA Polymerase (5 U/μl)	1.25 U/50 μl	0.25 μl
Total Volume	-	50 μl

Reagent	Final Concentration	Volume
Double-distilled water or Milli-Q water	-	(36.75-x) μl
10X PCR Buffer (with Mg2+)	1 X	5 μl
dNTP (2.5 mM each)	0.2 mM each	4 μl
Template DNA	10 pg - 1 μg	X μl
Primer Mix (10 μM each)	0.8 μM	4 μl
Taq DNA Polymerase (5 U/μl)	1.25 U/50 μl	0.25 μl
Total Volume	-	50 μl

2. The recommended PCR reaction program is as follows:

Step	Temperature	Time	Cycles
Preliminary Denaturation	94℃	3 min	1
Denaturation	94℃	30 sec
Annealing	55℃	30 sec	30
Extension	72℃	1 min
Final Extension	72℃	10 min	1

Note:

If multiple similar PCR reactions are to be performed, a master mix containing water, buffer, dNTPs, and Taq enzyme can be prepared in bulk and then aliquoted into individual PCR tubes. Primers may also be included in the master mix depending on circumstances. The PCR reaction conditions including temperature, time, and cycle numbers should be adjusted according to different templates, primers, PCR product lengths, and GC content.
Typically, a final primer concentration of 0.2 μM provides good detection results, but this can be adjusted between 0.1–1.0 μM if necessary. Increasing the primer concentration may improve amplification efficiency, while decreasing it can reduce non-specific reactions.
The recommended template DNA amounts for a 50 µl reaction volume are as follows: mammalian genomic DNA: 0.1–1 µg; E. coli genomic DNA: 10–100 ng; plasmid DNA: 0.1–10 ng. Excessive template DNA may lead to non-specific PCR products.
Before starting the reaction, gently pipette to mix or briefly vortex to ensure homogeneity, then centrifuge at room temperature for a few seconds to collect liquid at the bottom of the tube. If the PCR machine does not have a heated lid, add a drop of mineral oil to the top of the reaction mixture.
The extension time should be set based on the length of the PCR product, typically allowing 1 minute per kb. For initial PCR reactions, it is recommended to set the number of cycles to 35 to maximize the chance of obtaining the desired PCR product.

Download the product instruction manual

Q：PCR产物非常少或没有特异性条带是什么原因呢?

A：引物设计不佳是PCR过程中最常见的问题。请选择适当的引物设计软件进行引物设计，注意引物的GC含量、二级结构、二聚体、退火温度、长度、特异性等方面的问题。在加入酶切位点等的引物中，一定要注意加入酶切位点等后整条引物的GC含量、二级结构、二聚体、退火温度、长度、特异性等方面的问题。在原有引物效果不佳的情况并且阳性对照引物可以正常工作的情况下，可以考虑更换引物。

Q：待扩增片段GC含量偏高怎样做？

A：GC含量较高的情况下PCR会变得相对比较困难，此时可以使用适合扩增高GC含量DNA片段的GC-rich buffer，并相应地根据GC-rich buffer的要求或说明调整PCR反应参数的设置。

Q：PCR Mix可以减少体系么？

A：可以，使用的引物、模板等比减少使用。

Q：PCR Mix有没有3’-5’的外切酶活性？PCR Mix扩增产物类型？

A:没有3’-5’的外切酶活性，产物带有3’-A。可用于TA克隆。