Algae total RNA - low quality?

cement_head · 05-27-2015, 08:29 AM

Hi,

I'm sure I already know what forum members will say - but I'm having issues isolating high quality total RNA from algae (BA trace attached). I think this sample is poor quality and degraded.

I used UltraSpec reagent.

I am going to now try TriZOL. Any other suggestions?

Thanks,
Andor

nucacidhunter · 05-27-2015, 09:35 PM

I would pay some attention to sample collection and storage as well. No extraction method or reagent would result in good quality RNA if it is already degraded.

dhatziioanou · 05-28-2015, 12:30 AM

Snap freeze in liquid nitrogen when you collect it then put it straight into trizol and grind it with Qiagens fancy bead machine or manually grind it with a mortar and pestle on dry ice while keeping it super frozen by adding liquid nitrogen to keep it super cool and from there put it straight into trizol and sonicate it (be aware that if any liquid nitrogen get into the tube with trizol and you close it it will burst open with a good bang).

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05-27-2015, 09:35 PM	#2
nucacidhunter Jafar Jabbari Location: Melbourne Join Date: Jan 2013 Posts: 1,238	I would pay some attention to sample collection and storage as well. No extraction method or reagent would result in good quality RNA if it is already degraded.

05-28-2015, 12:30 AM	#3
dhatziioanou Junior Member Location: Greece Join Date: Aug 2012 Posts: 8	Snap freeze in liquid nitrogen when you collect it then put it straight into trizol and grind it with Qiagens fancy bead machine or manually grind it with a mortar and pestle on dry ice while keeping it super frozen by adding liquid nitrogen to keep it super cool and from there put it straight into trizol and sonicate it (be aware that if any liquid nitrogen get into the tube with trizol and you close it it will burst open with a good bang).