Appendix I: Solutions For Nucleic Acid Blotting - Thermo Scientific Shake n Stack 6240 Operating And Maintenance Manual

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