Thermo Scientific Shake 'n Stack 6244, Operating And Maintenance Manual

Page: 82 / 92

© Thermo
Scientific
, May 2003. Issue 7 29
APPENDIX I
SOLUTIONS FOR NUCLEIC ACID BLOTTING & HYBRIDISATION
PROCEDURES
1. 20 x SSC (1 litre)
NaCl 175g
Sodium citrate 88g
Dissolve in 800ml of H
2
O and adjust pH to 7.0 with NaOH solution. Adjust the
volume to 1 litre with H
2
O and sterilise by autoclaving.
2. 20 x SSPE (1 litre)
NaCl 175g
NaH
2
PO
4
H
2
O 27.6g
Na
2
EDTA 7.4g
Dissolve in 800ml of H
2
0 and adjust pH to 7.4 with NaOH solution. Adjust the
volume to 1 litre with H
2
O and sterilise by autoclaving.
3. 100 x Denhardt’s Reagent
Ficoll 2g
Polyvinyl pyrollidone 2g
Bovine serum albumin (Fraction V) 2g
Dissolve in 100ml H
2
O, filter and store in aliquots at -20
°
C.
4. Salmon Sperm DNA
Dissolve the DNA at a concentration of 1mg/ml in sterile H
2
O, sonicate to a length
of 200-500bp, and then stand in a boiling water bath for 10 minutes. Aliquot and
store at -20
°
C.
5. Denaturing Solution
NaCl 1.5M
NaOH 0.5M
Sterilise by autoclaving.