Hi all,
So I have been doing high-throughput RNA extractions on xylem from Eucalyptus hybrids (urophylla x grandis). My extraction combines a custom CTAB protocol with silica column binding and purification of the RNA. In a single column format this protocol works great (>50ng/ul, 260/230 between 1.8 and 2.0 etc), but it isn't going great in a 96-well plate, so I am busy optimizing. Generally my yield is a bit low due to the nature of the samples, but I have never had this banding phenomenon circled in blue. The nanodrop readings of these samples aren't fantastic either (below 10ng/ul and 260/230 below 1.00). I have worked with samples before that have organic contamination, but have never seen this banding on a gel before. Any suggestions as to what might be causing this? For context this is a normal 1% agarose gel stained with ethidium bromide and I also treat the gel with bleach beforehand to get rid of any RNases in the agarose.
Any suggestions would be highly appreciated!
Hello Nadine Kleinhans
What is your loading dye made of? looks like its concentration is too high.
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