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RNA related questions
Answers to questions commonly asked regarding RNA services
What is the best way to isolate RNA?
It is the experience of our facility and the Nucleic Acid facility that using one of the RNeasy kits from Qiagen is the best way to prepare RNA. Some labs still prefer to initially isolate RNA using Trizol but it has been our experience, for microarray experiments and Real Time PCR, it is best to further purify RNA isolated with Trizol using RNeasy.
What is the best way to measure the concentration of RNA samples?
Our facility houses a NanoDrop spectrophotometer which can be used free of charge by facility customers to measure RNA concentration. The instrument requires only 1.5 ul of sample and has a detection range of 1.5 ng/ul to 3000 ng/ul. We have found that this spectrophotometer is much more accurate than many of the cuvette based instruments currently being used on campus.
What is the best way to determine RNA sample quality?
RNA sample quality is crucial for successful microarray analysis. We encourage all of our customers to have their RNA samples run on our Agilent Bioanalyzer which will provide an objective measure of sample quality. The Bioanalyzer allows a much better assessment of RNA quality than conventional gel electrophoresis and consumes much less sample. The Bioanalyzer requires only 1 ul of sample at a concentration of 50-500ng/ul.
How many samples can be run on a single Agilent Bioanalyzer chip and what volume and concentration do you require?
A Bioanalyzer chip can run up to 12 samples. While the Bioanalyzer requires only 1 ul of sample at a concentration of 50-500 ng/ul we request that you provide us with 5 ul of sample in this concentration range. We will use 1.5 ul on the NanoDrop to confirm that the sample concentration is within the instrument range and we will use 1 ul for the actual Bioanalyzer run. If requested we will return all unused samples.
I have a large number of samples that I need to isolate nucleic acid from, is there an easy way to do this?
Yes. You can use the Qiagen TissueLyser which is housed in our facility. This instrument allows the rapid simultaneous lysis of up to two 96-well plates of samples