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==Step 2: Probe purification with ethanol precipitation== *Add in required volume of 100% ethanol, 3M NaOAC and Glycoblue into tray {| border="1" align="center" | align="center" style="background:#f0f0f0;"|'''Content''' | align="center" style="background:#f0f0f0;"|'''Volume Needed''' | align="center" style="background:#f0f0f0;"|'''Volume''' |- |100% Ethanol||align="right"|2.5X Sample Volume||align="right"|8000uL |- |3M NaOAC||align="right"|0.1X Sample Volume||align="right"|320uL |- |Glycoblue||align="right"|1/300X Sample Volume||align="right"|10.7uL |- |Sample||align="right"|Sample Volume||align="right"|3200uL |- |Total Volume |colspan="2" align="right"|11530.7uL |- |} *Pool all 32 wells PCR product into tray using multichannel pipette(P300) *Mix all content by repeat pipetting using 10mL Pipette *Aliquot to 2 x 15mL tubes *Store in -80C for more than 20 mins *Turn on Centrifuge fix temperature at 4C *Centrifuge frozen 15mL tubes at 3000rpm for 20m at 4C *Remove all liquid '''CAUTION: DO NOT DISCARD BLUE PELLET AT THE BOTTOM OF THE TUBE''' *Add in 350uL cold 75% Ethanol transfer pellet to 1.5mL tube *Addin another 350uL 75% ethanol wash the 15ml tube and transfer to 1.5mL tube *Centrifuge at 10,000rpm for 5min at 4C *Remove all liquid, air dry sample for 5min or until samples dried out *Resuspend the DNA with 100uL RNAse free H2O
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