WebFeb 1, 2024 · 10× T4 DNA ligase buffer and 400 U/μL T4 DNA Ligase. 10. 20 mg/mL bovine serum albumin. 11. 20 mg/mL Proteinase K. 12. 5 M sodium chloride. 13. ... Mix at room temperature for 10 min (see Note 1). 5. Add 2.5 M glycine solution to a final concentration of 0.2 M (see Note 2). 6. WebHere we report microwave-induced specific cleavage, ligation, dephosphorylation, and phosphorylation of nucleic acids catalyzed by restriction endonucleases, T4 DNA ligase, T4 polynucleotide kinase, and calf intestinal alkaline phosphatase. The microwave-mediated method has dramatically reduced the reaction time to 20 to 50 s.
Microwave-mediated enzymatic modifications of DNA
WebJun 13, 2024 · Potential application of T4 DNA ligase in microbial mutagenesis was explored by mutating and screening PHB producing E. coli XLPHB strain. When applied to atmospheric and room temperature plasma (ARTP) microbial mutagenesis, large survival pool was obtained. Mutants available for subsequent screening for desirable features. WebNote: T4 DNA Ligase is unstable on ice for long periods. Therefore, Invitrogen recommends the enzyme be kept at -20 °C until within 5-10 minutes of use and returned … festool cxs ersatzakku
Tips for Maximizing Ligation Efficiencies NEB
WebNEB T4 DNA ligase buffer, 100 ul of 10% Triton X-100, 12 ul of 10 mg/ml BSA, and 1 ul of 2000 U/ul T4 DNA Ligase and incubating at room temperature for 4 hours. Samples were pelleted at 2500 G and resuspended in 432 ul water, 18 ul 20 mg/ml proteinase K, 50 ul 10% SDS, 46 ul 5M NaCl and incubated for 30 minutes at 55’C. WebThe Quick Ligation™ Kit enables ligation of cohesive end or blunt end DNA fragments in 5 minutes at room temperature. (25°C ) Fast - 5 minutes for cohesive or blunt ends Convenient - ligation performed at room temperature Flexible - suitable for all common ligation reactions WebNote: T4 DNA Ligase should be added last. The table shows a ligation using a molar ratio of 1:3 vector to insert for the indicated DNA sizes. ... For blunt ends or single base overhangs, incubate at 16°C overnight or room temperature for 2 hours. Heat inactivate at 65 degrees C for 10 minutes. 00:10:00. Start. Chill on ice and transform 1-5 ... hp india mail id