High sensitivity Southern blotting without radioactivity.

Detection of biotinylated DNA on a Southern blot using the North2South System and a cooled CCD imaging system. A Southern blot of biotinylated DNA samples from a DNA fingerprinting kit was detected using the North2South System and imaged for one minute using a cooled CCD camera system.

The Thermo Scientific North2South Hybridization and Detection Kit combines a novel enhanced luminol substrate for horseradish peroxidase (HRP) with optimized pre-made hybridization and blocking buffers to ensure consistent results and high sensitivity. Any nucleic acid blot can be hybridized with the appropriate biotinylated (biotin-labeled) oligonucleotide probe. After a brief membrane blocking step, the biotin label is detected with streptavidin- HRP and a 5-minute incubation with the chemiluminescent substrate. Results are easily recorded by brief exposure to X-ray film (1-15 minutes) or CCD camera.

Features

• Sensitivity equal to or greater than ³²P
• Much faster than DIG detection systems
• Amenable to nearly any DNA or RNA hybridization method
• Optimized hybridization and blocking buffers and protocol steps yield consistent results from experiment to experiment
• High level of light output and consistent low background allows for very short and multiple exposure times using cooled CCD camera systems or film
• Ready-to-use buffers and short process time makes switching from radioactive detection to chemiluminescent detection an easy transition, even for the novice user
• Specific plaques can be picked out of areas that are very dense with signal due to extremely low background

Related Resources and Products:

North2South Chemiluminescent Kit FAQ
LightShift Chemiluminescent EMSA Kit

References:

1. Adilakshmi, T. and Laine, R.O. (2002). J. Biol. Chem. 277(6), 4147-4151.
2. Bach, S., et al. (2002). Infect. Immun. 70(2), 988-992.
3. Borst, E-M., et al. (1999). J. Virol. 73(10), 8320-8329.
4. Haertel-Wiesmann, M., et al. (2000). J. Biol. Chem. 275(41), 32046-32051.