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* What do I need to provide to the platform to prepare a standard Takara SMART seq Ultra low input library?
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This protocol has been optimized for cDNA synthesis starting from 10 pg of total RNA. However, if
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your RNA sample is not limiting, we recommend that you start with more total RNA (up to 10 ng).
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Purified total RNA should be in nuclease-free water.
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1. **10 pg to 10 ng** of high quality total RNA in a maximum volume of **10µl**.
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1. If possible, a **separate aliquot of 2,5** µl of the same RNA preparation taken at the time of the preparation of the sample.
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1. The concentration of RNA in your sample and the 260/208 and 260/230 nm UV ratios (nanodrop)
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\--
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Before starting the library prep we will perform a QC (RIN score) on an Agilent BioAnalyser using the 2.5µl aliquot. The main sample is stored at -80°C until all RNA samples have passed QC.
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If the RIN-score is below 8 we will contact you and determine with you if you need/can provide a new sample, or if we proceed with the current sample.
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A RIN score below 8 can affect the quality of the obtained results. If no better sample can be obtained, an library prep adapted to degraded samples can be used. Please contact the genomics platform in this case |
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