Today's Procedure:
- After incubating roughly 24 hours at 56 C samples were sealed and shaken (lost some lysate due to poor seals on a few caps).
- Centrifuge at 5700 rpm for 20 second.
- Uncapped/Added 410 ul premade Buffer AL/EtOH solution.
- Capped with new caps. Shook for 20s.
- Centrifuge at 5700 rpm for 20 second.
- Placed a DNeasy 96 well column plate on the previously used S Block.
- Attempted to remove 600 ul of lysate from each sample
- Some had more some had less due to volume of tissue and loss of lysate. Most sample tubes appeared visually to be at roughly the same volume.
- Sealed plate with Airpore sheet.
- Centrifuged at 5700 rpm for 11 minutes.
- Decanted solute from S Block. Wiped with Paper towel.
- Removed Airpore tape.
- Added 500 ul Buffer AW1 to each sample
- Seal with Airpore tape.
- Centrifugee at 5700 rpm for 6 minutes.
- Removed Airpore tape.
- Added 510 ul Buffer AW2 to each sample.
- Centrifuged at 5700 rpm for 16 minutes. (Used no airpore tape per instructions)
- Moved DNeasy 96 well column plate to Elution Microtubes plate.
- Added 200 ul Buffer AE to each sample (Row A got ~100 ul)
- Sealed with Airpore tape.
- Incubated for 1 minute at room temperature.
- Centrifuged at 5700 rpm for 3 minutes.
- Removed Airpore Tape.
- Added 200 ul Buffer AE to each sample (Row A got 400 ul)
- Sealed with Airpore Tape
- Incubated for 1 minute at room temperature.
- Centrifuged at 5700 rpm for 3 minutes.
- Removed DNeasy column block from Elution Microtubes plate.
- Sealed Microtubes with rubber tops flipping which which side the openning tab was on for every column.
- Labelled sample with "O.lurida sample processed 2/12/2015. 09/14 Oyster, 10/14 Fidalgo Manchester. JH"
- Stored in the 4 C in 213.
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