PRT062607 (P505-15) HCl，137026

Competent Cell Preparation

and autoclave to sterilize.  2. TB solution:  *10mM PIPES  *15mM CaCl 2 (1.66g/L) *250mM KCl(18.6g/L) *Dissolve in nanopure water and adjust pH to 6.7 with KOH or HCl and then add MnCl 2  to 55mM

Yeast Cell Lysates

from stocks to give 50 fold dilution. Example: 20 ul of stock per 1ml buffer. Use IMMEDIATELY after adding protease inhibitors.   SUMEB BUFFER (1% SDS, 8M Urea, 10mM MOPS, pH 6.8, 10mM EDTA, 0.01% bromophenol blue) TO MAKE 100ml: 1.0ml 1M

In Situ HYBRIDIZATION TO TISSUE SECTIONS

DNase (1U/ml) to the 24ml reaction mixture and incubate 30' @ 37 degrees C. Stop the reaction: +60ml 10mM EDTA, 10mM Tris-HCl (pH 7.4), 0.2% SDS +1.67ml 5M NaCl +1.0ml 1M DTT Incubate 10-15' on ice Hydrolyze the probe (ideal