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==Step 8: Purify the product from the gel == * Switch gloves when using PCR hood 1. Prepare gel-shearing assemblies (2x) using non-stick 0.5ml and 1.6ml tubes. 2. Use G22 needle to punch a hole at the bottom of the 0.5 ml tube 3. Cut out 85-9 nt band. Divide into 2 equal pieces. Chop up each and place pieces into a gel-shearing assembly. 4. Centrifuge at top-speed > 12000 rpm for 3 min to shear gel. Move any remaining pieces to bigger tube. Remove the 0.5 ml tube 5. Add 400ul clean 1x TE buffer. 6. Vortex at 37 C for 1 hour 7. In PCR hood, transfer gel mix to Nanosep columns 8. Spin at top speed 15000 rpm for 3min 9. Transfer supernatant to 15 ml tube (sterile, RNA-ase, DNA-ase free). (For Set Y, supernatant were transfered to individual non-stick tubes)
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