Technical Overview

The basis of all blotting techniques (Southern,[1] North-ern[2] and Western blotting[3]) is the transfer of nucleic acids/proteins to a solid inert support/filter followed by hybridization with a specific labeled probe. The process involves depurination (for nucleic acids >5 kb) to allow a more efficient transfer and denaturation of double-stranded nucleic acids to provide a suitable target for the probe. Neutralization is required for nitrocellulose filters because nucleic acids are not retained on these supports at pH values above 9.0. Usually, transfer to the support is by capillary diffusion, although apparatus are available that enhance the transfer by using suction and fixation onto a nitrocellulose/noncharged nylon solid support by baking or UV cross-bonding, respectively. Hybridization of the specific probe is performed under optimal conditions, and washing to remove excess (nonspecific) hybridization is necessary before the degree of hybridization is determined.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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