AAV's now available from ADDGENE

These are exceptionally high quality with exceptional quality control and have been validated by Rothlab.

We launched the first 2 AAV on our website: 

We're hoping the release the non DIO versions and control vector next week!

Comments

  1. mrbossmanOctober 10, 2016 at 10:12 AM

    Is there a particular cre-expressing virus which works best with these?

    ReplyDelete
  2. zendudestOctober 10, 2016 at 10:14 AM

    Any Cre-expressing virus which has been validated should work fine

    ReplyDelete
  3. UnknownNovember 22, 2016 at 7:43 AM

    Hi, Dr. Roth! I had checked the virus titer on the addgene website (≥ 2×10¹² vg/mL), it seems it's less than previous UNC production (5.7x10¹² vg/mL). Have you tested the virus in vivo? Thank you!

    ReplyDelete
  4. zendudestNovember 22, 2016 at 7:58 AM

    Yes it works great.

    ReplyDelete
  5. UnknownJanuary 5, 2017 at 1:11 PM

    Dear Dr. Roth,
    If these viruses are injected into a Cre-line mouse, would they be expressed in the tagged neurons only? If not, do you recommend other viruses?
    Thank you

    ReplyDelete
    Replies
    1. UnknownJanuary 5, 2017 at 1:40 PM

      They mention these viruses are "Cre-dependent", so does this answer "yes" to my question?
      Also, there are different serotypes, do you recommend one versus the other?

      Delete
    2. zendudestJanuary 5, 2017 at 1:48 PM

      Yes. I typically recommend starting with AAV8

      Delete
    3. UnknownJanuary 5, 2017 at 1:53 PM

      Sorry to keep asking, but is there a specific reason for choosing this one over 2 for example? ( Note: I work on the olfactory bulb which is a pretty large area in case this changes things)

      Delete
    4. zendudestJanuary 6, 2017 at 11:14 AM

      No--we find it simply works nearly every time. You may wish to order several serotypes and simply test them side-by-side.

      Delete
  6. zendudestJanuary 5, 2017 at 1:17 PM

    The DIO versions will only be expressed in cells expressing Cre-recombinase--so yes

    ReplyDelete
  7. UnknownMarch 9, 2017 at 4:45 AM

    Hi
    Is someone familiar with the technique of Clozapine administration via drinking water or osmotic mini-pump's ?

    ReplyDelete

Post a Comment

Popular posts from this blog

CNO in drinking water

We frequently get requests regarding protocols for putting CNO in the drinking water for long-term studies. Here is our protocol: 1. Dissolve the CNO in a small volume of DMSO 2. Diluted the dissolved CNO in drinking water (e.g., 5mg/200ml) 3. Give mice with CNO drinking water  and protect from light using foil-wrapped bottles 4. The mice will receive 5mg/kg/day CNO (assume that mice weight 30g and consume 6ml water per day) 5. Prepare fresh daily. 6. A small amount of saccharine in the drinking water will mask the slightly bitter taste of CNO
Read more

Considerations for immunofluorescence for various DREADD constructs

Image
I receive frequent queries regarding the appropriate way to visualize DREADDs.  Below I show the three basic DREADD constructs: HA-tagged mCherry fusion proteins HA-tagged in a vector with IRES-mCitrene In unfixed tissue the mCherry and mCitrene can be visualized using the parameters from the table below or by going here . NOTE: only the mCherry fusion protein fluorescence will show that the DREADD receptor is expressed.  mCitrene simply will show which cells were transfected or transduced. For visualizing HA we have a detailed recommended protocol   the principle of which is below. I also show various secondary antibodies which might be used along with excitation/emission characteristics Here's what you might consider if you have an mCherry fusion protein: Finally, what if you are using one of the HA-hM3Dq-IRES-mCitrene or HA-hM4Di-IRES-mCitrene constructs. ...
Read more

How best to dissolve CNO ...it depends on what it is soluble in (and that depends on the particular polymorph)

I recently received the following question: " My question is with regard to the use of CNO in drinking water (in mouse studies). Your recipe calls for the addition of a small amount of DMSO to dissolve CNO at a concentration of 5 mg in 200 mL. As we and others routinely dissolve CNO in 0.9% saline (no DMSO) at concentrations up to four times this amount, I am curious whether the addition of DMSO to CNO solutions for H2O drinking is critical, or whether the properties of CNO permit it's direct dissolution in H2O at concentrations from 5-20 mg per 200 mL. There appears to be no difficulty dissolving such concentrations in H2O in my hands, but I would prefer to hear your perspective. Is there a published reference for the solubility profile of CNO in H2O or 0.9% saline?" It basically depends upon the source of the CNO.  CNO via the NIH RAID program (which is what we use) is not readily soluble in H20 while commercial CNO is.  Although we've verified that both source...
Read more