Etchevers:Notebook/STRA6 in eye development/2008/11/24

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Mouse tissue storage for ChIP trials

4x pregnant NMRI white mice of which removed embryos from 3. Keep the other one for later embryos if needed, perhaps for Stan for the later kidney (eg. next Monday).

Add in more details later, Nicolas' lab notebook has them.

  • Heather 12:13, 24 November 2008 (EST):

There were plenty of embryos, most of them viable and around the same stage, equivalent to E11 as expected. The nasal pits were present, the lens vesicle was closed, but no auditory hillocks or footplate (there was a side notch in the hindlimb bud, though, sometimes, meaning some of them may have varied to E11.5 - but never any pigmentation in eye. Theiler stage 18, perhaps 19.).

Dissected myself 31 eyes, Nicolas and Adeline had a hand at it too and dissected another 16 or so. I also dissected a few forebrains, essentially telencephalon and trying to get rhinencephalon, but it was after eye removal and I don't know how intact they really were.

We froze one tube with one brain, one tube with five brains, and four tubes with 10 eyes and one with eight. The eyes all had their periocular mesenchyme still.

These have been put away in the last two G rows and first 5 H rows (8 then 4x 10 eyes) of -80 yellow plastic box, first row.

  • They practiced sonication, getting best results between 5-7 pulses at 30W for first, 40W for subsequent pulses of 20 seconds, spaced for 20 seconds to cool off, in 15ml tubes with 400 μL. Decided on 4 because the majority of the chromatin smear was a little lower at 7 than at 5, where the biggest chunk was around 300 bp according to MW. Try to upload gel pictures now.