pK-matched running buffers for gel electrophoresis
US6582574B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Mar 28, 2000 |
| Grant date | Jun 24, 2003 |
| Priority date | — |
| Expiry date | Mar 28, 2020 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10T436/108331
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
pK-matched buffers, each containing two effective buffering components: one weak base and one weak acid which have similar pKa at 25° C. (within 0.3 pK units On agarose gels, the buffers in various concentrations were tested for separation of double-stranded DNA fragments with various DNA markers, agarose gel concentrations, and field strengths. Mobility was inversely proportional to the logarithm of molecular weight. The buffers provided high resolution without smearing at more dilute concentration than is possible with standard TAE (Tris/Acetate, pH 8.0) or TBE (Tris/Borate, pH 8.3) buffers. The buffers were also tested in 7M urea denaturing LongRanger™ sequencing gels and in non-denaturing polyacrylamide SSCP gels. The pK-matched buffers provide good separation and high resolution, at a broad range of potential pH values. In comparison to TAE and TBE, pK-matched buffers provide higher voltage and current stability, lower working concentration, more concentrated stock solutions, and lower current per unit voltage, resulting in less heat generation.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.