un691cs at genius.embnet.dkfz-heidelberg.de
Tue Nov 26 05:49:44 EST 1996
Well... one should never volunteer to give the recipee for
a deluxe hybridization buffer. But since I have been swamped with requests:
here it is !
*$* Slavemaster's High Stringency Hybridization Buffer:
100 ml ultrapure water (eg twice distilled, can be bought)
60 ml NaHPO4 buffer 1M, pH 7.4 (read Sambrook about how to prepare this)
250 ml formamide (eg Merck, good quality, treatment not necessary)
heat to 60C
add and stir:
7.3 g NaCl
0.2 g EDTA
35 g SDS
Adjust volume to 490 ml
let dissolve. cool to RT.
adjust pH to 7.1 - 7.3 with NaOH or HCl
adjust volume to 500 ml.
Herring sperm can be added (for concentration see Sambrook)
*$* The following buffer was honorary named after my technician, and is well
suited for probing with oligomers at 42C:
Uli's Low Stringency Hybridization Buffer
3 % SDS
4 x SSPE (Sambrook for 20 x SSPE)
5 mM EDTA
very viscous, heat prior to use.
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