Enzymatics--T4 DNA 连接酶 (快速)--L6030-HC-F
T4 DNA 连接酶 (快速)
摘要:适用于低浓度过夜连接模式
产品描述
T4 DNA连接酶催化双螺旋DNA或RNA中5′磷酸末端和3′羟基末端生成磷酸二酯键。该酶能够有效的连接平末端和粘性末端,并能够修复双螺旋DNA、RNA或DNA/RNA杂交链的单链缺口。
产品组分
Supplied In 10 mM Tris-HCl,50 mM KCl,1 mM DTT,0.1 mM EDTA,50% glycerol
pH 7.5 @ 25°C
Supplied With B6030 (10X T4 DNA Ligase Buffer),10X T4 DNA Ligase Buffer (B6030),500mM Tris-HCI,100 mM MgCl2,50 mM DTT,10 mM ATP,pH 7.6 @ 25°C
产品信息
货号 L6030-HC-F
浓度 600,000 U/ml
规格 26,000 U
产品特性
贮存温度 -25 to -15°C
纯度(SDS-PAGE) >99%
活性 300,000 U/mg
使用说明
建立反应体系:
Amount Description Final Concentration
2 µL 10X T4 DNA Ligation Buffer 1X
X µL Vector 1-10 ng/µL
X µL Insert 1-10 ng/µL
1 µL T4 DNA Ligase (120 U/ µL) 6 U/µL
X µL Type I Water N/A
20 µL Total Volume
Add all of above components to a clean reaction vessel, mix well by pipetting.Incubate at 25°C for 30 minutes.Immediately purify DNA using PCR clean-up column and elute in ~50 µL.
- OR–Immediately dilute (at least 1:10, but enough such that 0.1-10 ng of ligation product will be transformed) in TE or water. Transform 0.1-10 ng of ligation product into chemically or electrocompetent cell line that is compatible with vector.