Is 10X Single-Cell Sequencing Limited to Capturing the 3'UTR Region?
The 10X single-cell sequencing technology is based on GEM (Gel Bead in EMulsion) technology, where individual cells are co-encapsulated with barcoded gel beads in an oil-water emulsion. Each gel bead carries a unique barcode for labeling RNA molecules derived from the same cell. Upon cell lysis within the droplets, the released RNA hybridizes with the barcodes on the beads, forming RNA-barcode complexes. These complexes are subsequently reverse transcribed and amplified to generate cDNA, which is then subjected to sequencing.
10X single-cell sequencing is designed to capture mRNA transcripts, not limited to specific regions such as the 3'UTR. During cDNA synthesis, oligo-dT primers are employed to selectively anneal to the poly(A) tails of mRNA, initiating reverse transcription. Consequently, the captured sequences represent the 3' end of mRNAs, although complete full-length transcripts are not typically recovered. The method provides a representation of the 3' ends, including the poly(A) tail and adjacent sequences, but not the entire mRNA molecule from the 5' end to the 3' end.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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