Hybridization and Immobilization

Dilute immobilization probe into 10µM

Add 4.5µL of 1µM circular probe

Add 9µLof 10X 7.4 PBS buffer

Add 0.9µL of 10µM immobilization probe

Add 75.6µL RNase-free water

Vortex slightly and spin down

Heated to 90°C for 0h 0m 30s then cool down to 90Room temperature and keep for 0h 30m 0s

Take 22.5µL of 10mg/ml magnetic beads into a eppendorf

Vortex then spin down in Centrifuge 15000rpm

Carefully take the eppendorf onto the DynaMag and wait for 0h 1m 0s to collect the beads

Wash beads with 1X PBS for three times. Make sure that eppendorf should put on DynaMag when removing supernatant.

Add 90µL of hybridization solution into the eppendorf with dry beads

Incubate at 90Room temperature for 0h 30m 0s, then spindown in Centrifuge15000rpm

Carefully take the eppendorf onto the DynaMag and wait for 0h 1m 0s to collect the beads

Wash beads with 1X PBS for three times. Make sure that eppendorf should put on DynaMag when removing supernatant.

Preserve the functional beads in 90µL 1X PBS