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Sequence coverage (or depth) is the number of unique reads that include a given nucleotide in the reconstructed sequence. [1] [2] Deep sequencing refers to the general concept of aiming for high number of unique reads of each region of a sequence. [3] Physical coverage, the cumulative length of reads or read pairs expressed as a multiple of ...
Most high-throughput, next generation sequencing platforms produce shorter read lengths compared to Sanger sequencing.These new platforms are able to generate large quantities of data in short periods of time, but until methods were developed for de novo assembly of large genomes from short read sequences, Sanger sequencing remained the standard method of creating a reference genome. [10]
A unique barcode sequence used on the cell hashing antibody can be designed to be different from an antibody barcode present on the ADTs used in CITE-seq. This makes it possible to couple cell hashing with CITE-seq on a single sequencing run. [12] Cell hashing allows super-loading of the scRNA-seq platform, resulting in a lower cost of sequencing.
DNA sequencing is the process of determining the nucleotide order of a given DNA fragment. So far, most DNA sequencing has been performed using the chain termination method developed by Frederick Sanger. This technique uses sequence-specific termination of a DNA synthesis reaction using modified nucleotide substrates.
The sequence of nucleobases on a nucleic acid strand is translated by cell machinery into a sequence of amino acids making up a protein strand. Each group of three bases, called a codon , corresponds to a single amino acid, and there is a specific genetic code by which each possible combination of three bases corresponds to a specific amino acid.
For example, in 2018, a group of researchers used linked-read sequencing technology to sequence genetic information from a pregnant woman who was a carrier of Duchenne muscular dystrophy (DMD) mutation. [10] Linked-read sequencing allows them to identify the maternal haplotypes and determine the presence of the mutant alleles in the foetal DNA ...
Quality control assesses the quality of sequencing reads obtained from the sequencing technology (e.g. Illumina). It is the first step in sequence analysis to limit wrong conclusions due to poor quality data. The tools used at this stage depend on the sequencing platform.
Primer walking is a method to determine the sequence of DNA up to the 1.3–7.0 kb range whereas chromosome walking is used to produce the clones of already known sequences of the gene. [2] Too long fragments cannot be sequenced in a single sequence read using the chain termination method. This method works by dividing the long sequence into ...