What is 16S rRNA PCR?
Abstract. Background: Broad-range 16S ribosomal RNA (rRNA) gene polymerase chain reaction (PCR) is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection.
Why is 16S rRNA used in PCR?
16S rRNA Gene 16S rRNA genes have conserved and variable regions (Figure 8-3ai), where conserved areas reflect phylogenetic relationship among species (and are used as sites for PCR priming) and highly variable regions reflecting differences between species.
What is broad-range 16S rDNA PCR?
In summary, broad-range 16S rDNA PCR is a crucial adjunct to microbiological diagnostics as a second line when infection of a sterile site is highly suspected, but culture and qPCR for the most likely pathogens have been proven negative.
Which PCR is used to amplify 16S rRNA?
However, amplifying the 16S rRNA gene from a single bacterial cell using a primer set that corresponds to regions highly conserved among bacteria usually leads to the amplification of contaminating bacterial DNA. In the present study, we used Ex Taq in the single-cell PCR because it amplifies DNA highly efficiently.
What is the function of 16S rRNA?
The 16S rRNA is the central structural component of the bacterial and archaeal 30S ribosomal subunit and is required for the initiation of protein synthesis and the stabilization of correct codon-anticodon pairing in the A site of the ribosome during mRNA translation .
What is the function of 16s rRNA?
What is broad range PCR?
PCR targeting the gene encoding 16S ribosomal RNA (commonly named broad-range PCR or 16S PCR) has been used for 20 years as a polyvalent tool to study prokaryotes. Broad-range PCR was first used as a taxonomic tool, then in clinical microbiology. This changed bacterial taxonomy and allowed discovering many new species.
What is the 16S rDNA sequence used for?
16S rRNA or rDNA sequence analysis has become a major tool in the determination of relationships between bacteria, and it is widely used for identification purposes. The resolution offered by the 16S rRNA gene is not high enough to differentiate between closely related species of Psychrobacter, such as P.
Which primers were used for the 16S rRNA?
Since the 16S gene sequence is similar but not identical in different organisms, degenerate primers are used for 16S rRNA sequencing. A primer set is called degenerate when it is used as a mixture of oligonucleotide molecules that contain different nucleotides in defined positions.
Is 16S rRNA a primer?
Universal primers. The 16S rRNA gene is used for phylogenetic studies as it is highly conserved between different species of bacteria and archaea. Carl Woese (1977) pioneered this use of 16S rRNA. The most common primer pair was devised by Weisburg et al.