RNA Detection

4 Notes

1. The interaction of TO also adds thermal stability to the probe:
   DNA/RNA duplex, indicated by a 7–9C increase inTmfor
   13 nt probes.


2. The needle is reusable. For conducting multiple experiments,
   rinse with 70% ethanol and H 2 O whenever changing sample
   solution and at the beginning and the end of experiments.


3. Tools can be sterilized by autoclave or alternatives.


4. Fresh ice should be used and frequently changed during the
   experiment. Slushy ice can result in lower reviving rate, possibly
   due to better heat transfer efficiency.


5. We use a marker pen to mark 1 mm from the tip of the injection
   needle for depth indication.


6. The fast green dye should not spread too fast and too far into
   the neighboring area. Oligonucleotide probe solution may
   spread faster than plasmid solution because of lower viscosity.


7. Bleeding should be avoided but it happens during the proce-
   dure. We use a piece of Kimwipes to get rid of blood or any
   excessive liquid from the animal’s head before proceeding with
   the surgery.


8. A twitch of muscle on the head is normally observed due to the
   passing current. The twitching itself is harmless but it causes
   repositioning of the injecting needle and further damage to the
   brain tissue. We try to avoid such damage by retracting the
   needle/anode electrode to the surface of the skin immediately
   after electroporation.


9. Electroporated ECHO probes automatically accumulate in the
   nucleus. This phenomenon has been observed for in cultured
   cells and for other short linear oligonucleotide probes.

References

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2. Sutherland HG, Mumford GK, Newton K,
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   Bickmore WA (2001) Large-scale identification
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3. Misteli T (2007) Beyond the sequence: cellular
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4. Sleeman JE, Trinkle-Mulcahy L (2014)
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5. Mankodi A, Lin X, Blaxall BC, Swanson MS,
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270 Dan Ohtan Wang