NS5 Initial Sample Processing

Owned by Gregg Randolph
Last updated: May 11, 2021
2 min read

@Shannon Harris @Muhammad Saqib

  • Vortex and quick spin 96 tubes and load into a 96-position tube rack following order on the “Input” sheet

  • Label Tube Rack 5Initials# (First Plate from Abby Hoffman: 5AH1) and add this name to the first column of the “Working” sheet of the same document and the aliquot plate

  • 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 or Zymo Mock Community to fill out the last column and treat as samples. (ie. Abby’s second plate). Fill in info in Input sheet.

  • 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 “NovaSeq5”. 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.