5x quick ligase buffer  (New England Biolabs)

Journal: Journal of visualized experiments : JoVE

Article Title: Single-Cell Factor Localization on Chromatin using Ultra-Low Input Cleavage Under Targets and Release using Nuclease

doi: 10.3791/63536

Figure Lengend Snippet: 10. End repair, phosphorylation, adenylation

Article Snippet: Use the reaction conditions: 12 °C for 15 min, 37 °C for 15 min, 72 °C for 20 min, hold at 4 °C. table ft1 table-wrap mode="anchored" t5 Name Company Catalog Number Comments 1.5 mL clear microfuge tubes ThermoFisher Scientific 90410 1.5 mL tube magnetic rack ThermoFisher Scientific 12321D 1.5 mL tube-compatible cold centrifuge Eppendorf 5404000537 10 cm sterile tissue culture plates ThermoFisher Scientific 150464 10X T4 DNA Ligase buffer New England Biolabs B0202S 15 mL conical tubes VWR 89039-656 1X TE buffer ThermoFisher Scientific 12090015 200 μL PCR tubes Eppendorf 951010022 2X quick ligase buffer New England Biolabs M2200 Ligase Buffer 5X KAPA HiFi buffer Roche 7958889001 5X high fidelity PCR buffer 7-Amino-Actinomycin D (7-AAD) Fisher Scientific BDB559925 96-well magnetic rack ThermoFisher Scientific 12027 or 12331D 96-well plate VWR 82006-636 AMPure XP beads Beckman Coulter A63881 polystyrene-magnetite beads; Due to potential variability between AMPure XP bead lots, it is recommended that your AMPure bead solution be calibrated.

Techniques: Phospho-proteomics, Sterility, Cell Culture, Hood, Sequencing, Electrophoresis, Agarose Gel Electrophoresis, Purification, Illumina Sequencing, Control, Multiplex Assay, Negative Control, Saline, Transferring, Light Microscopy