SMART-Seq v4: Sorted Cells
Plate-based 1/4 Smart-Seq v4 - Sorted Cells
Our cell sorters have the capability to sort single cells into multi-well plates for subsequent preparation of single-cell sequencing libraries. Using a Mantis low volume liquid dispenser (Formulatrix) and a Biomek FXP Span-8 (Beckman Coulter) the Core will take isolated single cells prepare cDNA using the SMART-Seq v4 Ultra Low Input RNA Kit (Takara) and libraries using the Nextera XT DNA Library Prep Kit (Illumina).
For more information about the technology, please visit Takara SMART-Seq v4. Please go to the flow cytometry page to see a description of our cell sorters and contact information.
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Switching Mechanism at 5′ End of RNA Template (SMART) technology captures full-length transcripts from 1 - 250 cells per well
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Non-stranded full-length cDNAs, contains the 5' end of the mRNA transcript
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Alternatively spliced isoform detection & rare low abundance transcripts
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Tagmentation based library preparation
To submit sorted cells for 1/4 SMART-Seq v4 Library Preparation, please provide the following:
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Sort cells into 2 ul of Takara's SMART-Seq v4 Ultra Low-Input Kit Reaction Buffer (other buffers are not supported)
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Core can provide plates or tubes of the Reaction Buffer upon request
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The Reaction Buffer must be made fresh and kept on ice for no longer than 2 hours before flash-freezing sorted cells
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Contact our Flow Cytometry team to set up your sort with the Bauer Core (plate(s) of lysis buffer provided by the core)
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Other requirements:
- Samples must be arrayed in columns of a 96-well plate
- Samples and all appropriate Minilims requests must be submitted by the 1st of the month (or the Friday before as applicable)
- Turnaround time is 4 weeks from the 1st of the month
- Coordinate drop-off with Core Staff between 8 am – 4 pm