Parse Biosciences (Fixed) Single Cell Sequencing
The Parse Biosciences platform uses split-pool combinatorial barcoding to profile single-cell transcriptomes without the use of microfluidic instrumentation. The individual cells themselves function as compartments during multiple rounds of splitting, pooling, and ligation to generate different barcode combinations unique to each cell’s set of transcripts.
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Assay Requirements
- fixed cells or nuclei (using Parse Biosciences Fixation kit and following manufacturer’s protocol)
Fixed samples can be stored at -80 for up to 6 months prior to submission to the AGC for barcoding, library prep, and sequencing. Each prep contains enough barcodes to uniquely label up to one hundred thousand cells, which can be divided across 1-48 samples.
Frequently asked questions
Does the Parse Biosciences single cell solution target the 3' ends of transcript ends?
The Parse Biosciences platform uses both random hexamer and oligo (dT) primers during reverse transcription so full length transcripts (albeit with a slight 3′ bias) are targeted for sequencing.
How viable should my cells be?
For best results, cells should be 80-90% viable going into fixation.
How much sequencing do my samples need?
Parse Biosciences recommends a minimum of 20,000 reads/cell. Ultimately sequencing depth will depend on the transcriptional activity of your cells.