Size selection (Purification)
Yin-Tse Huang, Tsu-Chun Hung
Abstract
Size selection (Purification)
Steps
Calculate the amount of magnetic beads you need and DISTRIBUTE in a new 1.5 mL eppendorf tube ( Mix the bottle VIGOROUSLY to make sure it's homogeneous ).
For example, if you have 5 samples to clean up, distribute 49.5 μl in a new 1.5 mL eppendorf tube.
| A | B | C |
|---|---|---|
| Sample | Magnetic beads need | Magnetic beads need *1.1 |
| 1 | 9 μl | 9.9 μl |
| 2 | 18 μl | 19.8 μl |
| 3 | 27 μl | 29.7 μl |
| 4 | 36 μl | 39.6 μl |
| 5 | 45 μl | 49.5 μl |
| 6 | 54 μl | 59.4 μl |
| 7 | 63 μl | 69.3 μl |
| 8 | 72 μl | 79.2 μl |
In each 20µL sample, add 9µL magnetic beads in.
Mix sample and beads by gently flicking the tube for 5mins 0h 5m 0s
Transfer the tube to the 8-strip magnetic rack.
After all the magnetic beads are arrested, remove the supernatant.
Add 200µL 80% ethanol, flip the magnetic rack around to clean the pellet.
Wait until all the magnetic beads are arrested, remove the supernatant.
Repeat the step 5 .
Flash spin the tube, and put on the magnetic rack.
Remove ANY trace of liquid residuals using 10 μl pipette.
Add 10µL elution buffer.
Mix gently by flicking the tube and make sure all magnetic beads are dissolved in buffer, then flash spin down the sample.
Incubate the tube in PCR thermal cycler using " 37_incubation " program for 10min 0h 10m 0s .
Transfer the tube to the magnetic rack.
After all the magnetic beads are arrested, collect the 10µL supernatant to a 200 μl PCR tube or a 8-strip PCR tube.
The clean-up sample is now ready for 2' PCR.
