Description

PerfectHyb® hybridization solution contains an accelerator that improves the hybridization efficiency and background. The hybridization rate is accelerated and background hybridization minimized. The solution facilitates Southern and Northern blotting analysis.

Features

  • Efficient and sensitive RI and non-RI detection by Southern and Northern blotting analyses.
  • Reduces hybridization time from 16 to 1-2 hr.
  • Over night hybridization significantly increases the sensitivity on detection of low levels of expression mRNA by Northern blotting analysis.
  • Low viscosity and no generation of precipitates at room temperature.
  • It is not necessary to add salmon sperm DNA.
  • Hybridization and washing steps are performed at the same temperature.

Applications

  • Southern blotting
  • Northern blotting

Storage condition

Store at room temperature

Components

PerfectHyb® hybridization solution

Application data

Example 1. Comparison of efficiency for Northern blotting analysis.

Sample: HeLa cell total RNA 5µg
Target: Transferrin receptor gene
Probe: [32P]-labeled cDNA probe prepared by the random priming method

1: PerfectHyb®
2: Company A
3: Company B
4: Conventional method

The transferrin receptor mRNA was detected efficiently with low background on Northern blot analysis using PerfectHyb®.

Example 2. Comparison of the efficiency and specificity for detecting a gene polymorphism by Southern blot.

Sample: Genomic DNA Hoe III digested 5µg
Target: YNH24(D2S44)
Probe: [32P]-labeled probe

1: PerfectHyb®, 2hr
2: Company A, 2hr
3: Company B, 2hr

Polymorphic bands were efficiently detected with low background on Southern blot analysis using PerfectHyb®.

References

  • A.C. Mistry, S. Honda and S. Hirose, Structure, properties and enhanced expression of galactose-binding C-type lectins in mucous cells of gills from freshwater Japanese eels (Anguilla japonica). Biochem J. 360: 107-15 (2001)