Apr 26, 2023

Size selection (Purification)

DOI
https://dx.doi.org/10.17504/protocols.io.bp2l695eklqe/v1
  • 1Kaohsiung Medical University
Tsu-Chun Hung
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DOI: https://dx.doi.org/10.17504/protocols.io.bp2l695eklqe/v1
Protocol Citation: Tsu-Chun Hung 2023. Size selection (Purification). protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l695eklqe/v1
License: This is an open access  protocol  distributed under the terms of the  Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 26, 2023
Last Modified: September 13, 2023
Protocol  Integer ID: 81048
Keywords: purification, size selection
Abstract
Size selection (Purification)
Prepare 20 µL sample and 9 µL magnetic beads in 1.5 mL eppendorf tube.

ABCD
ComponentVolumeProportionNote
Sample20 μl1x
Magnetic beads9 μl0.45xBeaverBeads™ DNA Select Isolation

Mix sample and beads gently by flicking then flash spin the tube. Put on the regular rack for 5mins 00:05:00 .
Note
DON'T put the tube on magnetic rack during waiting in this step.

5m
Transfer the tube to the magnetic rack. After most of the magnetic beads attach to the wall, remove the supernant.
Add 300 µL 75% ethanol, flip whole magnetic rack around. Wait for 3 mins 00:03:00 and remove the supernant.

3m
Repeat step 4.
Quick spin the tube, and put on the magnetic rack. Remove superfluous solution with 10 μl pipette.

Note
Caution: DO NOT let the beads crack!

Add 10 µL elution buffer. Mix gently by flicking and flash spin the tube. Put on the regular rack for 10mins 00:10:00 .

Note
DON'T put the tube on magnetic rack during waiting in this step.

10m
Transfer the tube to the magnetic rack. After most of the magnetic beads attach to the wall, collect the supernant to 200 μl PCR tube or 8-strip PCR tube.
Ready for 2' PCR.