You need to work really cold. I spin down 50ml of culture then LN2 dip and freeze dry. I keep the sample cold as soon as I take it out of the freeze drier.
Put in 1-3 4-5mm stainless bearings and shake at 30hz for 1min in a frozen block.
Then straight into LN2 and put in warm lysis buffer (SDS or CTAB based)
I find that working as cold as possible cleaned up my microalgae rna heaps!
If you dont have freeze dryer and tissuelyser then you could try frezze thaw in cold lysis buffer with heaps of prot K
On Wednesday, January 9, 2019 at 11:13:29 PM UTC+10, najib abdellaoui wrote:
On Wednesday, January 9, 2019 at 11:13:29 PM UTC+10, najib abdellaoui wrote:
hello everyonei am working on chlorella vulgaris and trying to extract RNA from it using different methods:
- i tried to use liquid nitrogen combined with trizol,
- i tried to use plant kit from invitrogen,
however, i couldn't get good results when i run the RNA sample on gel. i couldn't get the 28 and 18S bands. i got just a smear at the bottom of the gel.can someone help me for the rna extractionthank you.
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