This protocol outlines how to use Zymo oligonucleotide purification columns.
| Item | Item article number | Distributor | Vendor |
|---|---|---|---|
| Zymo silica Oligo Clean & Concentrator kit | D4060 |
Nordic Biosite | Zymo Research |
| 100% or 95% ethanol | |||
| Elution buffer (e.g. nuclease-free water) |
If it is the first time you use the kit add 96 ml 100% ethanol (104 ml 95% ethanol) to the 24 ml DNA Wash Buffer concentrate (D4060)
or 192 ml 100% ethanol (208 ml of 95% ethanol) to the 48 ml DNA Wash Buffer concentrate (D4061).
The maximum capacity for the column is 5 µg. If you want to run more just run the same column severl times.
If the reaction is 30 µl add 20 µl of nuclease-free water to the 30 µl reaction mixture for a total of 50 µl.
Add 100 µl Oligo Binding Buffer to 50 µl sample.
Add 400 µl ethanol (95-100%) and mix well by pipetting.
Transfer the 450 µl sample to the Zymo spin column.
Add 750 µl DNA Wash Buffer to the column and centrifuge for 1 min at 10,000-16,000 x g.
Transfer the column to a new Eppendorf tube.
Add ≥ 6 µl elution buffer (water or TE) to the center of the column.
Centrifuge for 1 min at 10,000-16,000 x g.
The flow-through can immediately be used or stored frozen.
Measure the yield on NanoDrop and then elute further to desired stock concentration (usually 100 µM).