Does temperature affect post-hybridization washes?
Absolutely! As with hybridization, stringency is controlled by salt concentration, surfactant concentration, temperature and the duration of the wash. Typically, there are two stages of stringency washes. The first step is low stringency (friendly to all hybridized DNA on the membrane, but effective in removing unbound probe). The second step is usually performed at higher stringency (less salt, higher temperature), to remove incompletely hybridized DNA: — 2X SSC; 0.1% SDS at room temperature — 0.2X SSC; 0.
Absolutely! As with hybridization, stringency is controlled by salt concentration, surfactant concentration, temperature and the duration of the wash. Typically, there are two stages of stringency washes. The first step is low stringency (friendly to all hybridized DNA on the membrane, but effective in removing unbound probe).
