|
|
 |
|
Prices are in US dollars and valid only for US orders.
|
Description:
The NEBlot® Phototope® Kit is designed for the chemiluminescent detection of nucleic acids in standard Southern and Northern blotting applications. With the NEBlot Phototope Kit, biotin is incorporated into hybridization probes in two ways- first, by utilizing a unique biotinylated random octamer primer and second, by incorporating biotinylated dATP during polymerization. The biotinylated probes are hybridized to target DNA immobilized on a membrane, and the target DNA is detected by chemiluminescence using one of the Phototope Detection Kits (sold separately).
Advantages:- Stability: Biotinylated probes and reagents are stable for extended periods unlike 32P-labeled probes which are only stable for a few days.
- Speed: Only 40 minutes is required for the entire detection procedure. Exposure times of 5-10 minutes.
- Multiple Exposures: Light is emitted at a constant rate for days, so you can perform multiple exposures to optimize signal intensity. Re-exposure at a future date is achieved by simply adding more chemiluminescent reagent.
- Simple Reprobing: If you want to search the same membrane for a different set of fragments, simply strip the membrane and rehybridize with a different probe.
- Versatility: The Kit can be used for Northern blotting, Southern blotting, plaque or colony hybridizations.
- Simultaneous Detection of Biotinylated Molecular Weight Standards
- No Radioactivity
Kit Components:
5X Labeling Mix
Biotinylated dNTP Mixture
Klenow Fragment (3´→5´ exo–)
Pre-biotinylated Molecular Weight Markers
Unbiotinylated Control DNA
Storage Conditions
Storage Temperature:
-20°C
Notes
General notes:- Phototope® Kits require transfer of DNA from a gel to a solid support membrane. The kits were developed using Millipore Immobilon-S membranes; however, other nylon based membranes with a neutral or slight positive charge can be substituted. Nitrocellulose does not give acceptable results.
Southern Blot using the NEBlot Phototope Kit. Exposure time 10 minutes.
References
- Feinberg, A.P. and Vogelstein, B. (1983) Anal. Biochem., 132, 6-13.
- Feinberg, A.P. and Vogelstein, B. (1983) Anal. Biochem., 137, 266-267.
- Denhardt, D.T. (1966) Biochem. Biophys. Res. Commun., 23, 641.
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual (2nd Ed.), 9.31-9.62.
- Perry-O'Keefe, H. and Kissinger, C.M. (1989) F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl (Eds.), Current Protocols in Molecular Biology, pp. 3.19-3.19.8.
Reagents Sold Separately
5X Labeling Mix
Biotinylated dNTP Mixture
Klenow Fragment (3´→5´ exo–)
Companion Products
Phototope®-Star Detection Kit
|
