Amplified DNA was electrophoresed in a 1.0% (w/v) agarose gel in Tris-borate-EDTA buffer
, stained with ethidium bromide, and photographed.
The conditions for PFGE were 1% agarose gel in 0.5x Tris-borate-EDTA buffer
at 6 V [cm.sup.-1], with pulse times increasing linearly from 1 to 60 seconds within 24 hours.
Migration of the DNA fragments was performed in 0.5X Tris-Borate-EDTA buffer
in a 0.9% agarose gel at 17 [degrees] C.
For confirmatory electrophoretic analysis of PCR products, a 10-[micro]L aliquot of each reaction was run on a 2% gel in Tris-borate-EDTA buffer
Electrophoresis was performed with 1 X Tris-borate-EDTA buffer
at 200 V and 23 mA for 3-5 h at room temperature, stained with 0.5 mg/L ethidium bromide in 0.6X Trisborate-EDTA buffer for 30 min, and visualized under ultraviolet light.
Polyacrylamide (acrylamide:bis-acrylamide, 19:1, 40%) and 10x Tris-borate-EDTA buffer
were from Bio-Rad and BioWhitaker, respectively.
2, the HCV amplicon was easily detected at a migration time of ~13 min when a physical gel solution composed of 10 g/L HPMC and 10 [micro]mol/L EtBr in Tris-borate-EDTA buffer
at 23 [degrees]C was used.
The digested products were then loaded on 6% acrylamide:bisacrylamide (19:1, by weight) gels and electrophoresed in Tris-borate-EDTA buffer
. Electrophoresis was carried at 40 mA in 1X Tris-borate-EDTA buffer
A 20-[micro]L aliquot of each mixture was then combined with 4 [micro]L of 6 X Triple Dye Loading Buffer (FMC BioProducts) and was electrophoresed through a 50-cm Hydrolink-MDE gel (AT Biochem) containing 150 g/L urea in Tris-borate-EDTA buffer
(80.3 mmol/L Tris base, 48.5 mmol/L boric acid, 1.53 mmol/L [Na.sub.2]EDTA) for 15 h at 1000 V.
Electrophoresis of CFLP fragments was performed in 8% denaturing polyacrylamide gels, 15 cm X 17 cm X 0.5 mm, containing 0.5X Tris-borate-EDTA buffer
and 8 mol/L urea.
RNA samples were mixed with loading buffer (2.5 g/L bromphenol blue, 2.5 g/L xylene cyanol FF, 1 mmol/L EDTA, and 400 g/L sucrose in diethylpyrocarbonate-treated water) and applied to a 1.0% agarose gel, which was prepared in 1 x Tris-borate-EDTA buffer
. Electrophoresis was conducted in 1 x Tris-borate-EDTA buffer
containing 0.1 mg/L ethidium bromide, at a constant voltage of 100 V for ~30 min.
The capillary (length, 40 cm), coated with linear polyacrylamide (18), was rinsed with Tris-borate-EDTA buffer
(89 mmol/L Tris-borate buffer containing 1 mmol/L EDTA, pH 8.3) for 5 min and then filled with sieving medium consisting of 60 g/L SLPA in Tris-borate-EDTA buffer
containing 7 mol/L urea (pH 8.3).