Vortex and quick spin deep well plates.
Use the 50 ml Integra expandable spacing pipette to transfer 30 ul from each well into a transparent PCR plate. Label the plate as shown here.
If it is only a few shy of a full plate, add 30 ul TE to fill out the last column and treat as samples. (ie. Abby’s second plate)
Vortex and spin control oligo pool (16S and ITS coligo 0.01 pg/ul each AND 16S SG_GR and ITS SG_GR 0.03 pg/ul each. This should be in the left hand refrigerator. (MIGHT NEED TO DELETE THIS STEP IF ALL EXTRACTIONS HAD COLIGOS ADDED BEFORE EXTRACTION.)
Add 6 ul control oligo plate to the sample aliquot plate in a well to well fashion (A1 to A1, B1 to B1, . . .) This can be done using an 8 channel or the 96 channel. If you use the 8 channel, you can reuse the dome caps on the control plate as long as you avoid contamination. Return control plate. Label side opposite numbers of aliquot plate with an asterisk.(MIGHT NEED TO DELETE THIS STEP IF ALL EXTRACTIONS HAD COLIGOS ADDED BEFORE EXTRACTION.)Coligos added during extraction so do NOT add during sample processing.
Seal, vortex, and spin aliquot plate.
Make a new folder under DNA Quantification labeled “Alfalfa Buerkle Lab 16S/ITS”. Check concentration on Synergy HTX and save file on petalibrary as plate name. Add a circle around the asterisk when concentrations have been measured. Normalize plates on the Hamilton Nimbus to 10ng/uL.
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