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Shannon Harris Muhammad Saqib

  • Vortex and quick spin 96 tubes and load into a 96-position tube rack.

  • Label Tube Rack 4Initials# (First Plate from Abby Hoffman: 4AH1)

  • Use the 50 ml Integra expandable spacing pipette to transfer 30 ul from each tube into a transparent pcr plate. Label this plate the same as the tube rack.

  • 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. Send me a picture of the label to be certain. It should be the only plate in their marked 0.01 pg/ul coligo and 0.03 pg/ul synthgene.

  • 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.

  • Seal, vortex, and spin aliquot plate.

  • Check concentration on Synergy HTX and save file on petalibrary as plate name. Make a new folder under DNA Quantification labeled “NovaSeq4”. Add a circle around the asterisk when concentrations have been measured. This plate can go on the top shelf of the left hand refrigerator well sealed.

  • Scan tube barcodes into NovaSeq4 document. Scan in a column fashion; follow the well layout already in the document. Add our plate name to “Alternate Plate Name” for tubes add the same to “Plate Name” column. Add last name of researcher (Hoffman) to “Researcher” column.

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