Size selective DNA precipitation protocol

<html><h2>Solutions/reagents:</h2><ul type="circle"><li> <a name="DNA sample">DNA sample <tab><div style="margin-right: 600px;">(DNA to be separated (e.g. PCR reaction mixture))</div></a></li><li> <a name="PEG/MgCl2">PEG/MgCl2 <tab><div style="margin-right: 600px;">(30% (w/v) PEG 8000/30 mM MgCl2 (concentration of PEG 8000 can be varied to shift the size of the percipitated DNA. The concentration used here will remove DNA fragments with less than 300bp))</div></a></li><li> <a name="TE buffer">TE buffer <tab><div style="margin-right: 600px;">(10 mM TRIS-HCl, 1 mM EDTA, pH 8.0)</div></a></li><li>buffer of choice</li></ul><h2>Equipment:</h2><ul type="circle"><li>Centrifuge</li><li>Eppendorf tubes</li></ul><h2>Steps:</h2><ol><li>Measure out 50 µl of <a href="#DNA sample" >DNA sample</a> into Eppendorf tube (1). Add 150 µl of <a href="#TE buffer" >TE buffer</a>. Gently tap the mixture for a few secs. </li><li>Add 10 µl of <a href="#PEG/MgCl2" >PEG/MgCl2</a>. </li><li>Vortex the mixture for a few secs. </li><li>Centrifuge at a speed of 10000 Xg for 15 mins at room temperature, gently aspirate out the supernatant and discard it. </li><li>Carefully remove supernatant not to disturb the pellet, which will be invisible. </li><li>Add buffer of choice to pellet. Add appropriate volume of buffer. Dissolve the pellet in the solution. </li></ol></html>

Size selective DNA precipitation protocol

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

research

Tools