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en:ethanol_precipitation [2013/06/09 10:10] (current)
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 +
 +<select Language>​
 +bio:​エタノール沈殿|Japanese
 +bio:​en:​Ethanol precipitation|English
 +</​select>​
 +
 +
 +======Ethanol precipitation======
 +Precipitation of polynucleotide such as DNA and RNA with ethanol aqueous solution to purify the polynucleotide.
 +
 +
 +=====Procedure=====
 +
 +  -Add 1/10 volume of 3M AcONa.
 +  -Add 2x volume of cold 100% EtOH.
 +  -Incubate for 20 min at r.t.
 +  -Centrifuge at 4 ºC, 15000rpm for 10 - 15 min.
 +  -Remove EtOH by pippetting or decantation.
 +  -Add cold 70% EtOH aq. and mix.
 +  -Centrifuge for 5 - 10 min.
 +  -If necessary, dry by using centrifuging evaporator.
 +
 +
 +=====Purpose=====
 +  *To prepare dried DNA.
 +  *To remove excess salt from the solution.
 +  *To purify DNA from the solution after Ph/Chl protein precipitation.
 +===70% EtOH aq.===
 +|-|precipitates|solutes (removable)|
 +|nucleicacids|DNA,​ RNA|dNTP, NTP|
 +|salts|-|salts in the solution|
 +|solvents|-|organic solvents\\ (phenol and chloroform)|
 +===100% EtOH===
 +|-|precipitates|solutes (removable)|
 +|nucleicacids|DNA,​ RNA\\ dNTP, NTP(high conc.)|-|
 +|salts|salts in the solution\\ (high conc.)|-|
 +|solvents|-|organic solvents\\ (phenol and chloroform)|