Embryo manipulation/dissection: Difference between revisions
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imported>Virgilio (Created page with "'''In this section:''' :Agarose Plates, 1% (for use with trops or laevis) :Dehydrating embryos :Rehydrating Embryos(for embryos stored in 100% methanol) '''Agarose Plates, 1% ...") |
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Latest revision as of 09:00, 14 December 2011
In this section:
- Agarose Plates, 1% (for use with trops or laevis)
- Dehydrating embryos
- Rehydrating Embryos(for embryos stored in 100% methanol)
Agarose Plates, 1% (for use with trops or laevis)
1 g agarose powder + 100 ml 1/9 X MR without gent in 500 ml jar
- make no more than 300 ml (to prevent boil over)
Microwave on high 2 min/100 ml (w/ loose lid)
Swirl and check for solids
When completely melted, fill petri dish 1/4 full
Put in 4C for 5 min, or air dry 30 min
Lift out with tweezers
- Can use a mold in agar before it hardens, or cut out desired shape after hardens
Dehydrating embryos
Embryos can be placed directly into 100% methanol in small glass vials.
Embryos can be stored at -20 deg C.
Rehydrating Embryos(for embryos stored in 100% methanol)
Draw off methanol with vacuum
Rehydrate embryos stepwise through each wash, 5 minutes each, on rotating rack:
3:1 methanol:PBS
1:1 methanol:PBS
1:3 methanol:PBS
100% PBS, repeat 2 - 3 times