RNA concentration normalization wizard
|Reported by:||Nicklas Nordborg||Owned by:||Nicklas Nordborg|
Description (last modified by )
A new step that is performed immediately after RNA extraction and Nanodrop concentration measurements.
Instead of putting the extracts RNA in the freezer as is, an aliquot is taken and diluted to the concentration that we need in the current mRNA step. The remaining RNA is put in the freezer while the aliquot is stored temporarily since it should go into library preparation very soon.
In theory, if only such pre-diluted aliquots were used in the mRNA wizard the lab protocol would have only zeroes in the H20 column. The RNA dilution registration would not have to be used.
Also note that the mRNA wizard require that RNAQC items exists and have either RQS or RIN values. In a "real-time" scenario this will not be the case, so the wizard must work without those values.
Summary of changes
- At the end of the 'Create aliquots on Bioanalyzer/Caliper plates' wizard it should be possible to enter the 'RNA concentration normalization wizard'. The current RNA items are automatically selected (the wizard should support manual selection as well).
- RNA items with too low quantity are not normalized, but flagged with
- It should be possible to print a lab protocol with dilution volumes and place for comments.
- Registration is done as the last step and should create new items with '.a' as last part of the name and subtype=
RNANormalizedAliquot. The RNA items are annotated with
AutoProcessing=Disableso that the create mrna wizard doesn't select them.
- Labels for the tubes should be printed from the partion wizard together with the other labels.
- The mrna wizard should support selecting normalized aliquots. Mixing normalized with non-normalized RNA is not supported. Biggest difference is the underlying query used for automatic selection and that it should be possible to select items without any RQS or RIN value.
- The 'Lab protocols for mRNA and cDNA preparation' need to support the normalized items. No dilution is needed and storage box is temporary.
- 'RNA dilution registration' wizard doesn't have to be done on the plate.
- The rest of the downstreams wizards should be the same, but they probably have to be made aware of the extra item that may site between mRNA and RNA.
Change History (17)
comment:1 by , 9 years ago
|Summary:||RNA dilution wizard → RNA concentration normalization wizard|