BsaⅠ 限制性内切酶,specification,price,image-Bio-Equip in China

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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.