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1. Add 750ul Reagent B (50mM Tris [pH8], 10mM EDTA, 100mM NaCl, 1% [w/v] SDS) and 50ul 100mg/ml proteinase K to each spleen.
2. Incubate overnight at 56ºC.
3. Gently shake to dissociate tissue.
4. transfer 200ul to eppendorf tube.
5. Add 50ul 5M sodium perchlorate.
6. Incubate 25 mins 65ºC.
7. Add 200ul phenol/chloroform.
8. Mix by inversion for about 2 mins.
9. Spin 13000rpm for 2 mins.
10. Transfer aqueous phase (top layer) to new eppendorf.
14 Add 500ul ice cold 100% ethanol.
15. Spool DNA into 500ul TE 10:0.l (if no visible DNA see below).
16 Allow to resuspend at 4ºC overnight.
If no visible DNA:
1. Spin 15 mins 13000rpm at ºC.
2. Remove liquid.
3. Add 500ul 70% ethanol.
4. Spin 5 mins 13000rpm at 4ºC.
5. Remove liquid and vac dry.
6. Add 500ul TE 10:0.1.
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