How To Please Ron/Brain

  • AD blood cells from PrecisionMed? Talk to Brian?
  • Meet with Kyle about designing siRNA (email sent)
    • Make siRNA sequences in geneious
    • Send order to Brian
  • Picture of how low sensors come together

miRNA Sensors (Theory)

  • Determine which pairs of target sites we want
    • Find sequences (hopefully under 200 bp)
    • Send ultramer order to Brian
  • Plan new MAV vector
  • Plan high sensor construction (meet with Brian)

miRNA Sensors (Lab Work)

  • Golden Gate
    • Gather miRNA target sites from Jeremy
      • Nanodrop for concentrations ( ? )
    • Make all 6 single low sensors (in conjunction with antibody group)
  • Miniprep eBFP2

NOTES

Both LR's failed (mKate2 and L7ae)

Meeting w/ Jeremy tomorrow to talk about new MAV vector and high sensors

We need to Gibson in  eBFP2 before we can finish the sensors

 

siRNA MEETING WITH KYLE

mature (5' phospho) and sense (reverse complement)
copy format from 2013

send brian both sequences as a duplex (IDT has a duplex option), with all the modification
We NEED to Gibson in the eBFP2 control
each high sensor on its own transcript (not necessarily own plasmid)

QUESTIONS
what do the "r" and "m" stand for?
-rna and modification
where do the overhangs come from? why are they there?
-mature one has 2 u's (constant)

 

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