Linker Ligation (with T4 ) DNA Ligase In a microcentrifuge tube prepare a solution of blunt ended, dephosphorylated DNA (100-500ng) in TE buffer (5-7µl). Add 1-2µg of phosphorylated linkers in 5µl of TE buffer. Add: 10X ligation buffer 2µl, 50% PEG 4000 solution 2µ...
RNA ligation using T4 DNA ligase. In: Hartmann RK, Bindereif A, Schön A, Westhof E, editors. Handbook of RNA Biochemistry. Vol. 1. Weinheim, Germany: Wiley-VCH; 2005. pp. 36–52.Frilander M. J., Turunen J. J. (2005) RNA Ligation Using T4 DNA Ligase , Wiley-VCH, Hoboken, ...
We present cognate base pair selectivity in templateヾependent ligation by T4 DNA ligase using a hydrophobic unnatural base pair (UBP), Ds㏄a. T4 DNA ligase efficiently recognizes the Ds㏄a pairing at the conjugation position, and Ds excludes the noncognate pairings with the natural bases. Our...
If any of these factors are not optimized, the DNA ends of the insert and vector may fail to anneal frequently enough for ligase to seal the fragments together. Reaction components Here’s a recommended ligation reaction ...
Unit Definition:0.01 Weiss unit of T4 DNA Ligase is the amount of enzyme required to catalyze the ligation of greater than 95% of 1μg of λ/HindIII fragments at 16°C in 20 minutes. Protocols Complete Protocol T4 DNA Ligase, Blue/White Cloning Qualified ProtocolPDF(112 KB)– English ...
包含T4 DNA 连接酶,可确保增强性能。 性能指标 修饰酶亚型 DNA 酶 底物类型 DNA 每个试剂盒的反应次数 150 连接酶类型 T4 DNA ligase 适用于 Lambda arms Plasmid vectors 资料库 产品样本 Competent Cells: Difficult Cloning, Protein Expression, and General Cloning Comp Cells ...
V. GENERAL PROTOCOL USING QUICK DNA LIGATION KIT A. Ligation 1. Add your DNA insert and vector into one tube with a 3:1 molar ration, and then adjust volume to 5 μl.2. Add 5 μl of 2× Quick Ligation Buffer and mix.3. Add 1 μl of T 4 DNA Ligase and mix thoroughly.4. ...
Backbone and insert were ligated using T4 DNA ligase (New England BioLabs, #M0202). NEB 5-alpha competentEscherichia coli(high efficiency) cells (New England BioLabs, #C2987I) were transformed with the ligated plasmid. Transformed bacteria were plated on LB + Amp (100 µg/mL) agar...
A novel approach to building a limitless variety of mobile multiple-restriction DNA fragments termed “uni-linkers” and which are open only at one end, is described. The latter were used as “multi-stage” linkers in cloning experiments. The cloning of the cDNA for a carrot proline-rich ...
Rapid DNA Ligation Kit Protocol Summary of the Experimental Process: Figure 1: Process summary of 5min Universal Ligation Mix 1. Ligation Reaction Component Amount 1. Prepare the rection mix on ice: Component Volume Vector 0.03 pmol Fragment 0.03 – 0.3 pmol 2 x Universal Liga...