品牌咨询
联系方式
公司地址
苏州工业园区生物纳米园A4#216
联系电话
4000-520-616 / 18915418616
传真号码
0512-67156496
电子邮箱
info@ebiomall.com
公司网址
https://www.ebiomall.com

SMOBIO/[TF3000] G-HiFi™ DNA Polymerase, (1 U/μl, 100 U)/1 U/μl, 100 U/TF3000

价格
面议
货号:TF3000
浏览量:127
品牌:SMOBIO
服务
全国联保
正品保证
正规发票
签订合同
商品描述

 

Description 

The G-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase suitable for GC-rich templates that are difficult to amplify. The fidelity of G-HiFi™ DNA Polymerase is 70 times higher than that of Taq DNA polymerase. The high extension rate of G-HiFi™ DNA Polymerase is achieved by blending the DNA polymerase with an elongation enhancer. The optimized 5X G-HiFi™ Buffer includes special ingredients that suppress non-specific amplification as well as plateau effect produced by conventional PCR. With the optimized 5X G-HiFi™ Buffer, G-HiFi™ DNA Polymerase is capable to amplify most templates, such as longer targets (up to 40 kb from lambda DNA) and that contain GC-rich sequences. 

Features

  • 5’→3’ DNA polymerase activity 

  • 3’→5’ exonuclease (proofreading) activity 

  • High reaction rate: 7 seconds/kb 

  • High fidelity: 70 times higher than Taq polymerase 

  • Generates blunt end amplicons  

  • Vast elongation capability (up to 40 kb) 

  • Thermo-stable for more than 10 hrs at 95°C 

Storage

[TF3000] G-HiFi™ DNA Polymerase

-20°C for 24 months

Odoo - Sample 1 for three columns

Elongation capability

G-HiFi™ DNA Polymerase’s high processability enables reliable amplification of λDNA up to 40 kb in length (M: DM5100). 

Odoo - Sample 2 for three columns

Sensitivity

G-HiFi™ DNA Polymerase performs higher sensitivity for high GC content templates (GC: 71%) compare to high fidelity DNA Polymerase from Brand A (M: DM2000).

 

[TF3000] G-HiFi™ DNA Polymerase

Contents

Component

Volume

G-HiFi™ DNA Polymerase (1 U/μl)

100 μl

5X G-HiFi™ Buffer

1200 μl

dNTPs Mix (2 mM each)

600 μl

Storage Buffer

50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C. 

Storage

-20°C for 24 months 

 

[TF3000] G-HiFi™ DNA Polymerase

Manual

Manual_TF3000_G-HiFi™ DNA Polymerase

SDS

SDS_TF3000

 

 

Recommended PCR Condition

[TF3000] G-HiFi™ DNA Polymerase

Template

1 – 150 ng

Forward primer

0.1 – 0.5 µM*

Reverse primer

0.1 – 0.5 µM*

5X G-HiFi Buffer

10 µl

dNTPs (2 mM each)

5 µl

G-HiFi DNA Polymerase

0.5 – 1 unit**

H2O

to 50 µl

Total volume

50 µl

*When amplifying products  10 kb in length, use primers at a final concentration of 0.1 μM each.

** When amplifying products  2 kb in length, use 0.5 unit of G-HiFi DNA Polymerase.

 

Recommended PCR Program

For  10 kb products

Select primers with a Tm value of 55°C. 20- to 25-mer primers are suitable, or those greater than 25-mer in length may provide optimal results.

Steps

Temp.

Time

Cycles

Template denature

98°C

2 min

1

Denature

98°C

10 sec

25-40

Annealing

50-68°C

15 sec

Extension

68°C

10-30 sec/kb

Final extension

68°C

1 min

1

 *Optimal PCR condition varies according to primers’ thermodynamic properties.

For  10 kb products

Select primers with a Tm value of  65°C. 25- to 35-mer primers are suitable. Avoid high GC-content at the 3" end of each primer.

Steps

Temp.

Time

Cycles

Denature

98°C

10 sec

25-40

Extension

68°C

10-30 sec/kb

 

For GC-rich templates:

Steps

Temp.

Time

Cycles

Templatedenature

98°C

2 min

1

Denature

98°C

10 sec

25-40

Extension

68°C

10-30 sec/kb

AFEAP cloning: a precise and efficient method for large DNA sequence assembly

Fanli Zeng,1 Jinping Zang,1 Suhua Zhang,2 Zhimin Hao,1 Jingao Dong,corresponding author1 and Yibin Lincorresponding author3

BMC Biotechnol. 2017; 17: 81. Published online 2017 Nov 14. doi: 10.1186/s12896-017-0394-x

PMCID: PMC5686892

Odoo - Sample 2 for three columns

Gel electrophoresis

Staining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.   

Safe fluorescent dyes

  • [NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl

  • [DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl

  • [DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 ml

Blue-light illuminator

  • [VE0100] B-BOX™ Blue Light LED Epi-illuminator

Odoo - Sample 3 for three columns

Ligation

Blund-end PCR amplicons can directly ligate with PCR cloning vector.  

  • [CV1100] GetClone™ PCR Cloning Vector II, 20 Rxn

Odoo - Sample 3 for three columns

Transformation

Prepare competent cells with high efficiency and transform with time-saving protocol.

  • [CK1000] Champion™ E. coli Transformation Kit

Odoo - Sample 3 for three columns

Colony PCR

Analyze colonies with PCR master mix to save preparation time.  

  • [TP1100] ExcelTaq™ 5X PCR Master Mix, 200 Rxn

  • [TP1200] ExcelTaq™ 5X PCR Master Dye Mix, 200 Rxn

 

2004年,SMOBIO Technology,Inc.迅速出现在生物技术行业。SMOBIO Technology,Inc.首先采用独特的发酵技术,旨在生产高质量功能性食品成分。建立成功的分销网络后,我们将专注于高质量产品继续,我们开发了最先进的生物分子测试工具。我们的工具旨在通过提供以下内容来促进分子生物学和蛋白质科学研究:高质量DNA阶梯和蛋白质标记。这些创新产品已受到业界和学术界人士的高度评价。自2008年以来,SMOBIO Technology,Inc.已展示出非常可观的市场份额。客户满意的证据表明,SMOBIO Technology,Inc.竭尽全力生产高质量产品。我们日益增长的声誉高质量 由超过满意的客户产生的工具已成为周围最好的营销策略之一。