A typical RNA-seq run requires an input amount of at least 10 ng of total RNA. If for some reason, your available RNA is limited, the RNA can be amplified before being sequenced. Currently, we routinely apply the SMARTer Universal Low Input RNA Kit (Clontech) which yields up to 50 ng of cDNA from input as low as 200 pg of total RNA. These amounts can be obtained from only a few cells.
Advantages of Single Cell RNA-seq
- Low input amounts of RNA (200 pg) can be amplified to sufficient amounts for sequencing
Single Cell RNA-seq workflow
We initiate the process by performing an initial quality check of the total RNA provided by you. During this quality check the concentration and integrity of the RNA is determined. We only continue with samples that passed this initial quality check. If for some reason certain samples did not pass this initial check, we will contact you to discuss on how to proceed.
In the next steps, the RNA is amplified and transcribed to cDNA using the SMARTer Universal Low Input RNA Kit. From the amplified cDNA sequencing libraries are constructed, which are again quality checked. Finally, the libraries are sequenced on one of our Illumina sequencers. The technical quality of the sequencing run is monitored in real time.
Sequencing data can be transferred to you via the Illumina BaseSpace platform or our own server (FTP download).
Feel free to contact us with any questions.