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The sample throughput for 3' mRNA-seq library preparation differs per method but can allow up to 384 samples to be processed in plates, with options for automation. [2] [12] For methods where samples are pooled early in the workflow, consumable use and cost are further reduced. For instance, BRB-seq is up to 25 times cheaper than Illumina ...
A genomic library is a set of clones that together represents the entire genome of a given organism. The number of clones that constitute a genomic library depends on (1) the size of the genome in question and (2) the insert size tolerated by the particular cloning vector system. For most practical purposes, the tissue source of the genomic DNA ...
Schematic overview of the MERCURIUS BRB-seq workflow where up to 384 samples can be barcoded and multiplexed per kit.. Bulk RNA barcoding and sequencing (BRB-seq) is an ultra-high-throughput bulk 3' mRNA-seq technology that uses early-stage sample barcoding and unique molecular identifiers (UMIs) to allow the pooling of up to 384 samples in one tube early in the sequencing library preparation ...
A cDNA library is a combination of cloned cDNA (complementary DNA) fragments inserted into a collection of host cells, which constitute some portion of the transcriptome of the organism and are stored as a "library". cDNA is produced from fully transcribed mRNA found in the nucleus and therefore contains only the expressed genes of an organism.
When sequencing RNA other than mRNA, the library preparation is modified. The cellular RNA is selected based on the desired size range. For small RNA targets, such as miRNA, the RNA is isolated through size selection. This can be performed with a size exclusion gel, through size selection magnetic beads, or with a commercially developed kit.
To select mRNA, the RT is performed with a single-stranded sequence of deoxythymine (oligo dT) primer which bind specifically the poly(A) tail of mRNA molecules. Subsequently, the amplified cDNA library is used for sequencing. [64] So, the first step of the method is the single cell encapsulation and library preparation.