Team:UNIPV-Pavia/Protocols/Precipitation

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(→DNA precipitation with sodium acetate)
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     <font face="Pristina" size="50" ><b>Protocols</b></font>
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*We use it only if the miniprep did not give us a high yield of DNA
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*Add 1/10 DNA solution volume of sodium acetate 3 M
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*Add 1/10 digestion volume of sodium acetate 3 M
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*Add 2.5 DNA solution volume of absolute ethanol
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*Add 2.5 digestion volume of absolute ethanol
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*Freeze at -80°C for 30 min
*Freeze at -80°C for 30 min
*Centrifuge at 13000 rpm, 4°C for 20 min
*Centrifuge at 13000 rpm, 4°C for 20 min
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*Remove all supernatant with a pipette
*Remove all supernatant with a pipette
*Air dry pellet until ethanol is totally removed
*Air dry pellet until ethanol is totally removed
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*Elute with 5 µl ddH2O
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*Elute with 5-10 µl of ddH2O
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<tr bgcolor='yellow'><td>INGREDIENTS:
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*'''Sodium acetate'''
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*'''Absolute ethanol'''
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'''Sodium acetate'''<hr color='white' width='70%'>
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*'''Ethanol 70%'''
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'''Absolute ethanol'''<hr color='white' width='70%'>
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*'''ddH2O'''
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'''Ethanol 70%'''<hr color='white' width='70%'>
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*'''Previously cut plasmid'''
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'''ddH2O'''<hr color='white' width='70%'>
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'''DNA sample'''</font>
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Latest revision as of 10:32, 21 October 2009

EthanolPVanimation.gif



Protocols

DNA precipitation with sodium acetate

(estimated time: 1 hour)

  • Add 1/10 DNA solution volume of sodium acetate 3 M
  • Add 2.5 DNA solution volume of absolute ethanol
  • Freeze at -80°C for 30 min
  • Centrifuge at 13000 rpm, 4°C for 20 min
  • Decant supernatant
  • Add 50 µl of 70% ethanol
  • Centrifuge at 13000 rpm, 4°C for 20 min
  • Remove all supernatant with a pipette
  • Air dry pellet until ethanol is totally removed
  • Elute with 5-10 µl of ddH2O




INGREDIENTS:

Sodium acetate
Absolute ethanol
Ethanol 70%
ddH2O

DNA sample





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