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Bst 2.0 DNA Polymerase
Bst 2.0 DNA Polymerase displays improved amplification speed, yield and salt tolerance
Optimized for Loop-Mediated Isothermal DNA Amplification (LAMP)
Improved amplification reaction properties compared to Bst DNA Polymerase
Fast polymerization
Flexible reaction conditions including a higher salt tolerance and thermostability, as compared to wild-type Bst DNA Polymerase, Large Fragment (NEB #M0275)
Interested in glycerol-free or custom formulations? – contact our Custom Solutions Group
Bulk packaging may also be available and requested for large recurring orders. Learn More
产品信息
Bst 2.0 DNA Polymerase is an in silico designed homologue of Bacillus stearothermophilus DNA Polymerase I, Large Fragment (Bst DNA Polymerase, Large Fragment). Bst 2.0 DNA Polymerase contains 5´→3´ DNA polymerase activity and strong strand displacement activity but lacks 5´→3´ exonuclease activity. Bst 2.0 DNA Polymerase displays improved amplification speed, yield, salt tolerance and thermostability compared to wild-type Bst DNA Polymerase, Large Fragment.
产品来源
Bst 2.0 DNA Polymerase is prepared from an E. coli strain that expresses the Bst 2.0 DNA Polymerase protein from an inducible promoter.
One unit is defined as the amount of enzyme that will incorporate 25 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
反应条件
1X Isothermal Amplification Buffer Pack Incubate at 65°C
1X Isothermal Amplification Buffer Pack 20 mM Tris-HCl 10 mM (NH4)2SO4 50 mM KCl 2 mM MgSO4 0.1% Tween® 20 (pH 8.8 @ 25°C)
贮存溶液
10 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 0.1% Triton® X-100 pH 7.1 @ 25°C
热失活
80°C for 20 min
单位活性检测条件
50 mM KCl, 20 mM Tris- HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24 mer, 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 100 μM dTTP including [3H]-dTTP and 100 μg/ml BSA.
优势和特性
应用特性
Isothermal amplification (LAMP)
Applications requiring strand-displacement DNA synthesis
DNA sequencing through high GC regions
Rapid sequencing from nanogram amounts of DNA template
相关产品
相关产品
Bst DNA Polymerase, Large Fragment
Deoxynucleotide (dNTP) Solution Mix
Deoxynucleotide (dNTP) Solution Set
Isothermal Amplification Buffer Pack
Magnesium Sulfate (MgSO4) Solution
Bst 2.0 WarmStart DNA Polymerase
注意事项
Bst 2.0 DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
Reaction temperatures above 70°C are not recommended.
Bst 2.0 DNA Polymerase cannot be used for thermal cycle sequencing or PCR.
Specific reaction conditions will vary for different isothermal amplification applications. For best results, use 1X Isothermal Amplification Buffer.
操作说明、说明书 & 用法
操作说明
Loop-mediated Isothermal Amplification (LAMP)
Typical LAMP Protocol (M0537)
工具 & 资源
Web 工具
NEB LAMP Primer Design Tool
FAQs & 问题解决指南
FAQs
What is the difference between Bst DNA Polymerase, Large Fragment and Bst 2.0 DNA Polymerase?
Why would I use Bst 2.0 WarmStart® DNA Polymerase?
Can Bst DNA 2.0 Polymerase be used in other NEBuffers?
Can Bst 2.0 DNA Polymerase be used to blunt DNA?
Can Bst 2.0 DNA Polymerase be used to fill in 3′ overhangs?
Can Bst 2.0 DNA Polymerase be used to remove 5′ overhangs?
Can Bst 2.0 DNA Polymerase be heat inactivated?
Are NEB DNA Polymerases supplied with dNTPs?
What are the main causes of reaction failure using Bst 2.0 DNA Polymerase?
Does Bst 2.0 DNA Polymerase have an active 3’→5′ proofreading exonuclease?
Can Bst 2.0 DNA Polymerase be used in applications requiring thermal cycling?
Can Bst 2.0 DNA Polymerase initiate at a nick in the DNA?
Can Bst 2.0 DNA Polymerase be used in labeling reactions and partial fill in reactions?
Can Bst 2.0 DNA Polymerase be diluted?
When should Bst 2.0 DNA Polymerase be the enzyme of choice?
Can Bst 2.0 DNA Polymerase be used at temperatures other than 65°C?
Does Bst 2.0 DNA polymerase incorporate dUTP?
Does Bst 2.0 DNA polymerase have reverse transcriptase activity?
How do I use Antarctic Thermolabile UDG for carryover prevention in LAMP reactions?
Does NEB have a master mix for LAMP or RT-LAMP reactions?
What is LAMP and RT-LAMP?
问题解决指南
PCR Troubleshooting Guide
引用 & 技术文献
引用文献
产品引用文献查找工具
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更多引用文献
Poole, C.B., Ettwiller, L., Tanner, N.A., Evans, T.C., Jr., Wanji, S.,Carlow, C.K. (2015) Genome filtering for new DNA biomarkers of Loa loa infection suitable for loop-mediated isothermal amplification. PLoS One; 10(9), e0139286.. PubMedID: 23272258, DOI: 10.1371/journal.pntd.0001948
Sanchita Bhadra, Yu Sherry Jiang, Mia R Kumar, Reed F Johnson, Lisa E Hensley, Andrew D Ellington (2015) Real-Time Sequence-Validated Loop-Mediated Isothermal Amplification Assays for Detection of Middle East Respiratory Syndrome Coronavirus (MERS-CoV). PLoS One; 10, e0123126. PubMedID: 25856093, DOI: 10.1371/journal.pone.0123126
出版物
Tanner NA, Zhang Y, Evans TC Jr (2015) Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques; Feb, 59-68. DOI: 10.2144/000114253
Poole CB, Zhang Y, Evans TC Jr., Carlow CK (2012) Diagnosis of brugian filariasis by loop-mediated isothermal amplification. PLoS Negl Trop Dis; 6(12), e1948.. PubMedID: 2327225, DOI: 10.1371/journal.pntd.0001948
Tanner NA, Zhang Y, Evans TC Jr. (2012) Simultaneous multiple target detection in real-time loop-mediated isothermal amplification Biotechniques; Aug, 81-9. PubMedID: 23030060, DOI: 10.2144/0000113902