RNA injection solution

• 1. Transcribe mRNA from a fluorescent protein-encoding plasmid (mMessage mMachine kit, Ambion).

• 2. Determine the RNA concentration.

• 3. Prepare 3- to 5-µL aliquots. Store at –80°C.

  Repeated freeze–thaw cycles will degrade the RNA.

• 4. To prepare the injection solution, combine the following:

  • RNA, one thawed aliquot

  • KCl, 0.1 m final concentration

  • Phenol red, 2.5% (v/v) final concentration (optional)

  • Nuclease-free H₂O

    Amounts of RNA and H₂O will vary; the ideal amount of RNA to use must be determined empirically.

• © 2011 Cold Spring Harbor Laboratory Press