I am currently working on a project to analyze Koeleria macrantha accessions. Unfortunately, the lab that we utilize is on a separate campus and our sample preparation must be done the day before or hours in advance. The buffers that we are currently using are described in Bonos et al (2002) Crop Science 42:192-196 as a combination of Arumuganathan and Earle and Huff and Palazzo. (modified Galbraith buffer (chopping buffer) and phosphate buffered saline suspension solution. Stained with PI.
We are having difficulty getting well defined peaks and encounter significant amounts of debris (too long of centrifuge cycle?). I would consider using the Otto buffers described in Pecinka et al. (2006) Annals of Botany 98: 117-122 (and elsewhere) but analysis was conducted 10 minutes after preparation which is unfeasible with our arrangement.
Can prepared nuclei (stained or unstained) be stored over night? Do samples need to be analyzed within a certain amount of time.
I appreciate any thoughts.
Matt
pkron
Joined: 28 Sep 2007
Posts: 10
Posted:
Wed Nov 04, 2009 10:04 pm
My experience suggests that overnight storage is usually a bad idea, and in fact, samples should not be left staining for more than 2 hours. In some species, nuclei may begin to degrade in an even shorter time. However, I would be interested to know if other people have had good luck with overnight storage.
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