Pre-normalization changes due to NeoPrep

[Nicklas Nordborg]

The neoprep require only 100ng of input rna in 25-45µl. The current pre-normalization is hard-wired for the manual protocol which is optimized for 1.1µg in 50µl.

The pre-normalization need to be modified so that it also works with the NeoPrep. The required RNA quantity and volume should be selectable, with at least the following presets:

• The old (dUTP) method (1.1µg in 50µl)
• TruSeq manual method (0.5µg in 50µl)
• NeoPrep method (0.1µg in 45µl)

Due to the much lower quantity of RNA for NeoPrep the pre-normalization will need to support mixing a larger volume (to avoid volumes less than 1µl). This also means that the mRNA wizard need to be aware of this, since it currently assumes that everything is used from the pre-normalized aliquot.

Another issue with the mRNA wizard is that if there are existing pre-normalized RNA for the NeoPrep and the mRNA wizard is used with the intention of creating a manual TruSeq plate, the NeoPrep samples will be auto-selected and then flagged with NotEnoughRemainingQuantity. This makes it very difficult the various protocols unless all queues are processed so they are empty before filling with RNA for a different protocol. Clearly the current auto-select function in mRNA wizard need to be much smarter about what to auto-select. The wizard also need to know about the various library preparation protocols.

[johan]

TruSeq manual method is (0.5µg in 50µl)