Bst DNA Polymerase, Large fragment,UA BIOSCIENCE_specification/price/image

Product Detail
Company Profile

Product Specification

Synonyms	DNA polymerase I
Expression System	E.coli
Molecular Weight	67kDa (Reducing)
Purity	＞95% by SDS-PAGE
Tags & Cleavage sites	/
Tag	His Tag
Storage Buffer	50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、0.1% Triton® X-100、50% Glycerol、pH 7.1 @ 25°C
Stability & Storage	Store at -25 ~ -15℃ for 2 years
Reference	1. Nucleic Acids Research, 2000, 28(12):E63. 2. Chemical Communications, 2014, 50(28):3747-3749. 3. Current Protocols in Molecular Biology, 2014(Suppl.105):15.14.1-15.14.14.

Background

Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity. It can be use to isothermal amplification (LAMP), DNA sequencing through high GC regions and Rapid Sequencing from nanogram amounts of DNA template.

Components

50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、0.1% Triton® X-100、50% Glycerol、pH 7.1 @ 25°C
10* Reaction Buffer: 200 mM Tris-HCl、100 mM (NH4)2SO4、20mM MgSO4、100 mM KCl、1% Tween® 20、pH 8.8@25°C
Magnesium Sulfate (MgSO4) Solution：100mM MgSO4

Protocol

Incubate the following reaction at 65°C for 30–60 minutes

Component	Final Concentration
10X Isothermal Amplification Buffer II	1X (contains 2 mM MgSO4)
MgSO4 (100 mM)	6 mM (8 mM total)
dNTP Mix (10 mM)	1.4 mM each
FIP/BIP Primers (25X)	1.6 µM
F3/B3 Primers (25X)	0.2 µM
LoopF/B Primers (25X)	0.4 µM
Bst DNA Polymerase (8,000 U/ml)	320 U/ml
DNA or RNA Sample	> 10 copies or more
Nuclease-free Water	to 25 µl
Total Reaction Volume	25 µl

Guidelines

1.Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.

2.100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.

3.Reaction temperatures above 70°C are not recommended.

4.Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR

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 65°C

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.