Sam:LabNotes/Microbiome-new/2010-5-25

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Pobes design for specific capturing on unculturable mouse gut bacteria[edit]

Background[edit]

Process[edit]

  • Set up criteria on Primer-3KZ program (Length=18nt, Tm=55C, GC%=40~70).
  • Pick up candidate probes.
  • Check the specificity (Mega-blast to nt database). Check secondary structure.
  • Label with FAM on 5'-end.
  • Order them on IDT at 100nmole scale with standard de-salting purification.
File:Probe design 052510.doc

Discussion[edit]

  • I have concerns about the signal intensity. The unamplified DNA only, the binding of one probe won't provide enough "visialbe fluorescent emission"
    • I will try to label target with combined probes.
  • I also designed the Biotin-C12-probes for TSA-FISH, but they haven't been ordered yet, becaus the FAM-probe is much cheaper.