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Product Information

Product Name

Catalog No.

Size

Hieff UNICON® HotStart Taq DNA Polymerase, 5 U/μL

10729ES72

250 U

Hieff UNICON® HotStart Taq DNA Polymerase, 5 U/μL

10729ES76

500 U

Hieff UNICON® HotStart Taq DNA Polymerase, 5 U/μL

10729ES80

1000 U

Hieff UNICON® HotStart Taq DNA Polymerase, 5 U/μL

10729ES92

10000 U

Hieff UNICON® HotStart Taq DNA Polymerase, 5 U/μL

10729ES93

25000 U

Product Description

Hieff UNICON® HotStart Taq DNA Polymerase is a mixture of Hieff UNICON® Taq Antibody(Catalog #31301ES)and Hieff® Taq DNA Polymerase (Catalog #10101ES). Hieff UNICON® Taq Antibody had a high affinity for Hieff® Taq DNA Polymerase, and the activity of Hieff® Taq DNA Polymerase could still be blocked after being treated at 50℃ for 30 min. The product was completely inactivated by heating at pre denaturation temperature for 30 seconds, releasing DNA polymerase activity. The hot start Taq enzyme can effectively inhibit the amplification caused by non-specific annealing of primers.

Hieff UNICON® HotStart Taq DNA Polymerase is suitable for hot start PCR and qPCR. Hieff® Taq DNA Polymerase in this product is a heat stable recombinant DNA polymerase with high template affinity, which is very suitable for amplification of low copy template. The amplified product has 3 '- dA and can be easily cloned into T vector.

Package Information

Component Number

Components

10729ES72 (250 U)

10729ES76 (500 U)

10729ES80 (1000 U)

10729ES92 (10000 U)

10729ES93 (25000 U)

10729-A

HotStart Taq DNA Polymerase (5 U/μL)

50 μL

100 μL

200 μL

1 mL×2

1 mL×5

Product Application

Low copy gene amplification and genotype identification.

Unit Definition

Using the activated DNA of salmon sperm as template / primer, the activity was defined as one active unit (U) when 10 nmol of total nucleotide was ingested as acid insoluble substance at 74℃ for 30 min.

Quality Control

Closed Detection of DNA Polymerase Activity:In the Hieff UNICON® HotStart Taq Buffer (Mg2+ Plus),the activity release was less than 5% after incubation at 65℃ for 30 min.

DNA Polymerase Activity Release Detection:In the Hieff UNICON® HotStart Taq Buffer (Mg2+ Plus),the activity release was higher than 95% when heated at 95℃ for 30 sec.

Exonuclease Activity: In 20 μL reaction system, 10 U of this product and 0.6 μg of λ - HindIII were incubated at 74℃ for 1 h, and the electrophoresis band of DNA had no change.

Endonuclease Activity: In 20 μLreaction system, 10 U of the product and 0.6 μg of supercoiled pBR322 DNA were incubated at 74 ℃ for 1 h, and the DNA electrophoretic bands did not change.

Residual E.coli gDNA Test: The E.coil 16S rDNA gene was amplified by adding 2 U of the product into 50 μL system and using sterile ddH2O as template. After 35 cycles, the amplified products were subjected to 1% agarose gel electrophoresis and EB staining without amplification bands.

Shipping and Storage

Transport in ice packs,it can be stored at -20℃ for 2 years.

Protocol Reaction Setup(for 50 μL)

Component

Volume (μL)

Final Concentration

ddH2O

to 50

-

2×Taq Buffer(Contains Mg2+、dNTP)

25

DNA template

Appropriate amount

-

Forward primer (10 μM)

2

0.4 μM

Reverse primer (10 μM)

2

0.4 μM

Hieff UNICON® HotStart Taq DNA Polymerase (5 U/μL)

0.5

2.5 U/50 μL

【Notes】

1)Reagent use:Each component should be fully melted and mixed before use.

2)Concentration of polymerase:2.5 U/50 μL is recommended. It can be optimized between 1.25-5 U / 50 μL.

3)Final concentration of Mg2+:The final concentration of the system was 2 mM. If there is a special need, 25 mM MgCl2 can be used, with the interval of 0.2-0.5 mM.

4)Recommended use of different templates (50 μL reaction system):

Type of template

Template usage

Genomic DNA

50 ng-200 ng

Plasmid DNA

100 pg-20 ng

cDNA

1-5 μL (No more than 1/10 of the reaction system)

Thermal cycling protocol

Stage

Temperature

Time

Cycles

Pre-denaturation

95℃

1 min

1

Denaturation

95℃

10 sec

35-40

Annealing

60℃

20 sec

35-40

Extension

72℃

30 sec/kb

35-40

Final Extension

72℃

10 min

1

【Notes】

1)Annealing temperature and time: 50-60℃ is recommended. Low annealing temperature will lead to nonspecific amplification. The primers were designed according to the standard of fluorescent quantitative primers. The recommended annealing time is 20 sec, which can be adjusted within 10-30 sec

2)Extension temperature and time: 72℃ is recommended. The recommended time is 30 sec/kb.

3)Amplification products: Please store the PCR amplification products at - 20℃ to prevent DNA degradation.

4)Amplification procedure:This product is used for quantitative experiments. It can be amplified by two-step procedure, removing the extension step, and combining annealing and extension into one step. The corresponding time is usually 30 sec or set according to the instrument model. For example, Applied Biosystems 7300 needs more than 31 sec.

Notes

1)For your safety and health, please wear experimental clothes and disposable gloves.

15221439726

15221439726