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Then, the index 1 primer attaches, polymerizes the index 1 sequence, and is washed away. The strand forms a bridge again, and the 3' end of the DNA strand attaches to an oligo on the flow cell. The index 2 primer attaches, polymerizes the sequence, and is washed away. A polymerase sequences the complementary strand on top of the arched strand.
To determine the sequence, four types of reversible terminator bases (RT-bases) are added and non-incorporated nucleotides are washed away. A camera takes images of the fluorescently labeled nucleotides. Then the dye, along with the terminal 3' blocker, is chemically removed from the DNA, allowing for the next cycle to begin.
In genetics, a transcription terminator is a section of nucleic acid sequence that marks the end of a gene or operon in genomic DNA during transcription.This sequence mediates transcriptional termination by providing signals in the newly synthesized transcript RNA that trigger processes which release the transcript RNA from the transcriptional complex.
Sequencing by reversible terminator chemistry can be a four-colour cycle such as used by Illumina/Solexa, or a one-colour cycle such as used by Helicos BioSciences. Helicos BioSciences used “virtual Terminators”, which are unblocked terminators with a second nucleoside analogue that acts as an inhibitor.
Intrinsic terminator containing an RNA hairpin rich in guanine and cytosine as well as a region high in uracil. The two types of termination signals in bacteria are intrinsic and factor-dependent terminators. [4] Intrinsic termination occurs when a specific sequence on the growing RNA strand elicits detachment of RNA polymerase from the RNA-DNA ...
This sequencing method is based on sequencing by synthesis (SBS) principle and the use of reversible dye-terminator that enables the identification of each single nucleotid. In order to read the transcript sequences on one end, and the barcode and UMI on the other end, paired-end sequencing readers are required. [67]
An alternative and mutually exclusive form involves bases in the 3' segment of helix 1 with those in the 5' region of helix 5 to form a structure termed the anti-terminator form. [ 1 ] [ 2 ] [ 3 ] When SAM is unbound, the anti-terminator sequence sequesters the terminator sequence so the terminator is unable to form, allowing the polymerase to ...
Antitermination in lambda is induced by two quite distinct mechanisms. The first is the result of interaction between lambda N protein and its targets in the early phage transcripts, and the second is the result of an interaction between the lambda Q protein and its target in the late phage promoter.