Etchevers:Notebook/Genomics of hNCC/2009/04/24
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Spoke with Candice on Monday when she came back from vacation.
The chicken NCC vastly prefer Matrigel to collagen I, and are still growing after their thaw 10 days ago. They can be passed in 1/2 diluted trypsin in PBS (confirm this). They were on T25 and then trypsined into 2 ampules for freezer again (if I understand my notes right).
I hope this hasn't timed out.
R992b CCNt XX, R1113 CCNt, R1062 CCNc (the CCNt were Phox2b+), R1066, and R1033 ( I think these are here?) are underway.
Candice and Yves will transfect Phox2b into some cell type so as to have a + control for IHC next week, as there is BG fluorescence but RT-PCR says no transcripts. Will also extract these + cells as the PCR is tricky.
Teratoma cells - more and more adherent.
Don't forget the HIS for TMEM216 and for HIPK2. ---
Yesterday: The pCAGGS-GFP vector is in the -20°C freezer
The rest of the samples that arrived today are at -80°C (C15 R682 XY 71 mg, and C15 R700 XY 47 mg), as well as kidney from EFP0600137 14 sa. This latter is quite large and we may need to smash it in two. Perhaps place the agate morter and pestle in the -80°C freezer.
I labeled some new freezer boxes I to IV, 0 to 9 in two dimensions and, until such time as we must set up a proper database, I want to make a spreadsheet for my colleagues and myself on Googledocs to manage the four hundred samples that could be stored therein.
There were 750K cells in the ampule of R1113 p10 cells (CCNt) and, I worry, about 3000K yeast. Lots of suspicious little bright things in the cell count grid.
R837 55 was AK2-A in PBT, some limb cartilage+? liver -
58 was AK2-A in PBS, liver -, weaker 57 was AK2-P in PBT, not the same liver mostly - but cells _, cartilage of vertebrae +, weak CNS, inner ear +/- and some muscle cells ++ 59 was AK2-P in PBS same but a bit weaker 56 no primary, not anything, even the inner ear. White.
18883-1 slide 36 no primary, liver negative, kidney some faint in tubules, placenta not in the parenchyme but villosities ++++ (AP not inhibited? not the same?)
slide 35 AK2-A PBT same as above, except more kidney tubules and the ureter + slide 33 AK2-A PBS same slide 34 AK2-P +++ liver, +++ kidney, ++ in placental parenchyme as well as BG of villosities.
Don't forget to note the embedding of rabbit corneas.
I cut them out post-fixation of the entire eye, in which formalin or PFA was injected after death and enucleation. They had sejourned about 24h in fix, were rinsed, and brought to 70% EtOH. I changed, then brought down to Florence for sectioning (requested 41 slides of each of 3 blocks, HE of slides 1, 21, 41.)
L368 is now 090833 L433 is now 090834 L447 is now 090835.
Stephane's notebook has details of injections and the rest.