5× dna loading buffer, blue  (Meridian Bioscience)

Journal: Methods in enzymology

Article Title: Reconstituting the 4-Strand DNA Strand Exchange

doi: 10.1016/bs.mie.2017.11.013

Figure Lengend Snippet: Analysis of the DNA substrates, intermediates, and gapped DNA product by electrophoresis in a 1.4% agarose gel. Lane 1: 1-kb DNA ladder. Lane 2, 3, and 5: purified circular ssDNA pBS II SK (+) (100 ng), 2065 bp dsDNA (125 ng), and gapped DNA (125 ng), respectively. Lane 4: DNA products of the annealing reaction (415 ng).

Article Snippet: Buffers and Reagents list-behavior=unordered prefix-word= mark-type=disc max-label-size=0 NanoPure water Xho I restriction endonucleases (NEB) Alw NI restriction endonucleases (NEB) Bovine serum albumin (BSA), 20 mg/mL (NEB, cat# B9000S) Ethanol 200 proof TAE buffer: 40 m M Tris, 20 m M acetic acid, 1 m M EDTA, pH 8.0 TE buffer: 10 m M Tris–HCl, 1 m M EDTA, pH 8.0 Agarose, Type I-A, Low-EEO (Sigma cat# A0169) for analytical electrophoresis Agarose, Certified Molecular Biology (Bio-Rad cat# 161-3101) for purification of dsDNA and gapped DNA molecules by electroelution 1-kb DNA ladder Molecular Weight Marker 10 × DNA loading buffer for agarose gels (0.25% bromophenol blue, 50% glycerol, 1 m M EDTA, pH 8.0) Ethidium bromide staining solution, 2 μg/mL in water 1-Butanol ≥99.4% ACS reagent (Sigma cat# 360465) 10 × annealing buffer: 250 m M Tris-acetate, pH 7.5, 500 m M NaCl Formamide ≥99.5% (Sigma cat# F9037) 2.1.3.

Techniques: Electrophoresis, Agarose Gel Electrophoresis, Purification