BsaⅠ 限制性内切酶,UA BIOSCIENCE_specification/price/image

Product Detail
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Product Specification

Species	Bacillus stearothermophilus
Synonyms	BsaⅠ 限制性内切酶
Expression System	E.coli
Molecular Weight	64.9kDa (Reducing)
Purity	>95% by SDS-PAGE
Conjugation	Unconjugated
Tag	His Tag
Physical Appearance	Liquid
Storage Buffer	10 mM Tris-HCl、300 mM NaCl、1 mM DTT、0.1 mM EDTA、500 µg/ml BSA、50% Glycerol （pH 7.4 @ 25°C）
Stability & Storage	Store at -25 ~ -15℃ for 2 years
Reference	[1] Zhu Z, Samuelson J C, Zhou J, et al.Engineering Strand-specific DNA Nicking Enzymes from the Type IIS Restriction Endonucleases BsaI, BsmBI, and BsmAI[J].Journal of Molecular Biology, 2004, 337(3):573-583. [2] Lee J H, Won H J, Oh E S,etal. Golden Gate Cloning-Compatible DNA Replicon/2A-Mediated Polycistronic Vectors for Plants[J]. Frontiers in Plant Science, 2020, 11.

Background

BsaI is a Type IIs restriction enzyme that can recognize non-palindromic sequences and cut outside of the recognition sequence. It is commonly used for Golden Gate assembly and enzymatic cleavage of plasmids to prepare linear DNA fragments with poly (A/T/G/C) endings and obtain specific sticky ends.

Recognition site:

5'-GGTCTC(N)1↓-3'

3'-CCAGAG(N)5↑-5'

Components

Storage Solution: 20 U/μl BsaⅠ、10 mM Tris-HCl、300 mM NaCl、1 mM DTT、0.1 mM EDTA、500 µg/ml BSA、50% Glycerol（pH 7.4 @ 25°C）

10*Reaction Buffer：500 mM Potassium Acetate、200 mM Tris-acetate、100 mM Magnesium Acetate、1mg/ml BSA（pH 7.4 @ 25°C）

Protocol

This step is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled up according to experimental needs.

1）Add the following components in sequence

Components	Volume
Plasmid DNA	1μg DNA
10*Reaction Buffer	5μl
BsaⅠ (20 U/μl)	1μl
RNase-free ddH2O	Up to 50μl

2）Incubate at 37°C 1h

3）DNA linearization is complete, and subsequent experiments can be performed.

Guidelines

1. star activity may result from a glycerol concentration of >5%; 2. Please avoid repeated freeze-thaw cycles.

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

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.