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Primers

The 16S rRNA gene is crucial for prokaryotic phylogeny and taxonomy, featuring conserved and variable regions that aid in primer design for amplification. The document lists various primers for amplifying different hypervariable regions of the 16S gene and provides details on the rpoB gene primers as well. It highlights the significance of these genes in microbial identification and diversity studies.

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Noelia Delgado guevaa
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33 views4 pages

Primers

The 16S rRNA gene is crucial for prokaryotic phylogeny and taxonomy, featuring conserved and variable regions that aid in primer design for amplification. The document lists various primers for amplifying different hypervariable regions of the 16S gene and provides details on the rpoB gene primers as well. It highlights the significance of these genes in microbial identification and diversity studies.

Uploaded by

Noelia Delgado guevaa
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Secuenciación 16S

The 16S rRNA gene is the most widely used macromolecule in prokary-otic
phylogeny and taxonomy investigations (del Rosario-Rodicio &del Carmen-
Mendoza, 2004). This gene alternates areas common toall microorganisms where
the sequence is known (conserved) withregions that change over time (variable).
The 10 conserved zones (C1–C10) are useful for designing primers that permit the
amplification ofthe hypervariable zones. Conversely, the nine variable regions (V1–
V9) provide the most helpful information for phylogeny and taxonomystudies.The
scientific community has established that the 16S rRNA genehas an approximate
average length of 1500 base pairs (bps) (delRosario-Rodicio & del Carmen-
Mendoza, 2004).

Tabla 1. Primers para amplificar diferentes regiones variables del gen 16S

Region Codigo
Gen Primers Primers
V primers

F:AGAGTTTGA F:
TYMTGGCTCA TGTAAAACGACGGCCAGTAGAGTTTGATYMTGGCTCAG
G ; R:GCT R: CAGGAAACAGCTATGACGCTGCCTCCCGTAGGAGT
V1-V2 27F/338R GCCTCCCGTA
GGAGT

F: AGA GTT F:
16S TGA TYM TGG TGTAAAACGACGGCCAGTAGAGTTTGATYMTGGCTCAG
CTC AG ; R:
ATT ACC GCG R: CAGGAAACAGCTATGACATTACCGCGGCTGCTGG
GCT GCT GG
V1-V3 27F/534R
F: CCT ACG F: TGTAAAACGACGGCCAGTCCTACGGGNGGCWGCAG
GGN GGC
WGC ;R:
AG ;R:GAC TAC CAGGAAACAGCTATGACGACTACHVGGGTATCTAATCC
V3-V4 341F/785R HVG GGT ATC
TAA TCC

F: GTG CCA F:
GCM GCC GCG TGTAAAACGACGGCCAGTGTGCCAGCMGCCGCGGTAA ;
GTA A ;R: GGA R: CAGGAAACAGCTATGACGGACTACHVGGGTWTCTAAT
CTA CHV GGG
V4 515F/806R TWT CTA AT

F: GTG CCA F:
GCM GCC GCG TGTAAAACGACGGCCAGTGTGCCAGCMGCCGCGGTAA ;
GTA A ;R: GAA R: CAGGAAACAGCTATGACGAATTAAACCACATGCTC
TTA AAC CAC
V4-V5 515F/944R ATG CTC

F: GAA TTG F:
ACG GGG GCC TGTAAAACGACGGCCAGTGAATTGACGGGGGCCCGCAC
CGC ACA AAG ;R:
AG ;R: CGG CAGGAAACAGCTATGACCGGTGTGTACAAGGCCCGGGA
V6-V8 939F/1378R TGT GTA CAA ACG
GGC CCG GGA
ACG

F: CAA CGA F: TGTAAAACGACGGCCAGTCAACGAGCGCAACCCT ;R:


GCG CAA CCC CAGGAAACAGCTATGACTACGGYTACCTTGTTACGACTT
T ;R:TAC GGY
TAC CTT GTT
1115F/
V7-V9 ACG ACT T
1492R

SECUENCIACIÓN gen rpoB


Tamaño
del
Primers Primers cola M13 Referencia
amplicón
(bp)
F: Mollet, C., Drancourt,
TGTAAAACGACGGCCAG M., & Raoult, D. (1997).
F:
TCAGTTCCGCGTTGGCC rpoB sequence analysis
CAGTTCCGCGTTGG
TG ; R: as a novel basis for
CCTG ;R: 687
CAGGAAACAGCTATGAC bacterial
CGCAACGGCCTGAC
CGCAACGGCCTGACGTT identification. Molecular
GTTGCAT
GCAT microbiology, 26(5),
1005-1011.
F: F:
GCCTTGCCAGCCCG TGTAAAACGACGGCCAG
CTCAGTCCGTGCAC TGCCTTGCCAGCCCGCT
CCCACCcaytayggnmg CAGTCCGTGCACCCCAC
; R: Ccaytayggnmg ; R:
GCCTCCCTCGCGCC CAGGAAACAGCTATGAC 387
ATCAGCCCACGGCC GCCTCCCTCGCGCCATC A Comparison
TGCckytgcatrtt AGCCCACGGCCTGCckyt of rpoB and 16S rRNA
gcatrtt as Markers in
Pyrosequencing Studies
of Bacterial Diversity

Otros primers

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