16S ribosomal sequencing
Targeted sequencing of the 16S rRNA gene for cost‑effective bacterial and archaeal community profiling at scale.
Pricing
Pricing preserved from the legacy page. Request a quote for current totals, turnaround, and volume discounts.
MR DNA has a huge collection on different 16s , ITS, 18s, COI, functional genes (e.g. nif, nir ) other taxonomic markers such as rpoB and we can adapt any primers to an amplicon sequencing technique using the bTEFAP(R) system.
NEW: Long read amplicons (e.g. near full 16s ~1400bp, ITS1-4, 18s ~1700bp or other custom amplicon diversity (700-2500bp) ( usually minimum 10 samples or $350 library fee applies) long reads = better taxonomic differentiation right?
1-10 samples $80/assay (small project fee of $150 added .. this fee is not per sample but per assay)
$80 for 20-50 assays,
$75 for 50-100 assays,
$70 for 100-150 assays,
$65 for > 150 assays.
MR DNA now accepts any size project (projects with < 10 samples do have a $325 indexing fee)
Academic and Government Pricing
$85 per sample for 5,000 sequences per assay,
$115 per sample for 10,000 sequences per assay
$180 per sample for 20,000 sequences per assay
$300 per sample for 40,000 sequences per assay
**Custom assays from 700bp - 3000bp or larger of course have a barcoding fee that is typically $15/barcode
DNA extraction starts at $40
--RNA Extraction starting between $50 - $70 / sample with RNA there is a fee for cDNA formation (depending on service requested and sample type submitted)
Microbial 16s, ITS, 18s, functional gene sequencing is helping to improve our understanding of human health, disease, and microbial evolution. The human body contains trillions of cells with a variety of microbes that play a critical role in human health and disease, but the area of mechanism remains a mystery. Microbes are not only present in the human body; they are everywhere e.g. human or animal guts, homes, plants, oceans, and soil. Microbiome sequencing was revolutionized in 2006 when Dr Scot E Dowd invented the barcoding, tagging of amplicons for next generation sequencing, from there scientists are now investigating this vast microbial world and microbiome has become a household term. Vast numbers of studies have now been published in the last 5-10 years examining the microbial communities that exist inside our bodies and how these microbiomes can be influenced by the environment. The microbiome of the human gut can be rapidly and accurately cataloged by amplicon or shotgun metagenomic sequencing via the Illumina NovaSeq 6000 System. Fecal samples, which have an abundant amount of microbes present (making it ideal to extract microbial DNA for genome sequencing), are a great candidate to help determine what is happening in the human GI tract. With NGS systems like the Illumina NovaSeq 6000, the scientific community is now able to generate gigabytes of data per sample. Many researchers today are excited about the value of microbiome sequencing and a number of clinicians believe it will become a routine part of health care, much like a blood draw.
Overview
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Platforms
Applications
- Microbiome Diversity
- Soil Ecology
- Food Science
- The 16s gene contains both highly conserved regions as well as hypervariable regions. The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice. The presence of the hypervariable regions affords researchers the ability to differentiate between closely related genera or species detected in their samples.
- The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice.
- The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice.
- Soil Ecology
- Food Science
- Sequencing Applications 16s Ribosomal Sequencing Genome Sequencing Metagenomics Microbial Sequencing Exome Sequencing Transcriptome Sequencing Amplicon Sequencing (bTEFAP®) Illumina AmpliSeq Ion AmpliSeq Cancer Genomics De Novo Sequencing Targeted Sequencing Small RNA Sequencing Single Cell RNA Sequencing Microbiome Diversity Bisulfite Sequencing
Methods and options
- Illumina NovaSeq 6000
- Illumina HiSeq 2500
- Illumina MiSeq
- Illumina AmpliSeq
- Contact us to discuss additional Illumina specific primer sets and pricing options
- The 16s gene contains both highly conserved regions as well as hypervariable regions. The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice. The presence of the hypervariable regions affords researchers the ability to differentiate between closely related genera or species detected in their samples.
- The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice.
- The overall size of the 16s rRNA gene is relatively short. ~1500bp. While sequencing the entire 16s gene is difficult due to read length restrictions of many NGS platforms, sequencing one or more hypervariable regions is relatively quick and affordable. Two of our most requested assays for 16s rRNA sequencing are 27F-519R (V1-V3 region) and 515F-806R (V4 region). For questions regarding pricing feel free to contact us or visit our 16 ribosomal sequencing page.
- Two of our most requested assays for 16s rRNA sequencing are 27F-519R (V1-V3 region) and 515F-806R (V4 region).
- The presence of the highly conserved regions allow researchers to design primer pairs that will accurately and reliably amplify the 16s hypervariable region of their choice.
What you receive
- Microbiome Diversity
- Sequencing Applications 16s Ribosomal Sequencing Genome Sequencing Metagenomics Microbial Sequencing Exome Sequencing Transcriptome Sequencing Amplicon Sequencing (bTEFAP®) Illumina AmpliSeq Ion AmpliSeq Cancer Genomics De Novo Sequencing Targeted Sequencing Small RNA Sequencing Single Cell RNA Sequencing Microbiome Diversity Bisulfite Sequencing
Long-form details (preserved for SEO)
Illumina NovaSeq 6000
Illumina HiSeq 2500
Illumina MiSeq
PacBio Sequel
Ion S5 XL / PGM
SeqStudio Genetic Analyzer
Sequencing Services
16s Ribosomal Sequencing
Genome Sequencing
Metagenomics
Microbial Sequencing
Exome Sequencing
Transcriptome Sequencing
Amplicon Sequencing (bTEFAP®)
Illumina AmpliSeq
Ion AmpliSeq
Cancer Genomics
De Novo Sequencing
Targeted Sequencing
Small RNA Sequencing
Single Cell RNA Sequencing
Microbiome Diversity
Bisulfite Sequencing
COVID-19
Bioinformatics
Development
Research
Free Software
Applications
Resources
Frequently Asked
Questions
Blog
Contact
16s Ribosomal Sequencing
Sequencing Services
16s rRNA Sequencing
Genome Sequencing
Metagenome Sequencing
Microbial Sequencing
Exome Sequencing
Transcriptome Sequencing
Amplicon Sequencing (bTEFAP ® )
Illumina AmpliSeq
Cancer Genomics
DeNovo Sequencing
Targeted Sequencing
Small RNA Sequencing
Single Cell RNA Sequencing
Microbiome Diversity
Bisulfite Sequencing
MR DNA specializes in any type of amplicon sequencing.
From 16s, 18s, ITS, functional genes to any type of custom primer amplicon assay.
We have an extensive inhouse assay (primer) collection for 16s sequencing, 18s sequencing, ITS sequencing, functional genes such as nirS, nifH, dsr, pufM, nosZ, HMC, and many more. if you have a custom assay we inexpensively set it up for you.
Amplicon Sequencing Prices (illumina).
MR DNA has all the major sequencing platforms and we have a large selection of amplicon sequencing programs .. Everything can be customized to the needs of the customer, only limited by the capabilities of the sequencing technologies.
Any amplicons (bTEFAP® services) such as 16s, 18s, ITS, functional or custom assays.. if you have an amplicon with custom primers we can help sequence it.
Illumina miseq and hiseq amplicons
examples for any of our hundreds of inhouse assays
2x300bp PE illumina 20,000 sequence diversity assays
additional discounts for very large projects also
PAC BIO SEQUEL:
Sequel long read amplicon sequencing for 16s, 18s, ITS and custom amplicons
DNA Extraction / RNA Extraction Price:
16s rRNA Sequencing Primer List
Updated Earth Microbiome Project (EMP) 16s v4
515F — GTGYCAGCMGCCGCGGTAA
806R — GGACTACNVGGGTWTCTAAT
Original Earth Microbiome Project (EMP) 16s v4
515F — GTGCCAGCMGCCGCGGTAA
806R — GGACTACHVGGGTWTCTAAT
Other Common 16s rRNA Primers
515F — GTGYCAGCMGCCGCGGTAA
926R — CCGYCAATTYMTTTRAGTTT
909R — CCCCGYCAATTCMTTTRAGT
archaea 349F — GYGCASCAGKCGMGAAW
A344F — AYGGGGYGCASCAGGSG
archaea 806R — GGACTACVSGGGTATCTAAT
arch21F — TTCCGGTTGATCCYGCCGGA
arch519R — TTACCGCGGCGGCTG
arch1059R — GCCATGCACCWCCTCT
arc344F — ACGGGGYGCAGCAGGCGCGA
arch915R — GTGCTCCCCCGCCAATTCCT
27F — AGRGTTTGATCMTGGCTCAG
519Rmod — GTNTTACNGCGGCKGCTG
519Rmodbio — GWATTACCGCGGCKGCTG
1492R — GGGTTACCTTGTTACGACTT
338R — AGTGCTGCCTCCCGTAGGAGT
28F — GAGTTTGATCNTGGCTCAG
519R — GTNTTACNGCGGCKGCTG
341F — CCTACGGGNGGCWGCAG
785R — GACTACHVGGGTATCTAATCC
805R — GACTACNVGGGTATCTAATCC
799F — ACCMGGATTAGATACCCKG
1193R — CRTCCMCACCTTCCTC
a799wF — AMCVGGATTAGATACCCBG
new1193R — ACGTCATCCCCACCTTCC
16com1F — CAGCAGCCGCGGTAATAC
16com2R — CCGTCAATTCCTTTGAGTTT
926F — AAACTYAAAKGAATTGACGG
1394R — ACGGGCGGTGTGTRC
Tx9F — GGATTAGAWACCCBGGTAGTC
1391R — GACGGGCRGTGWGTRCA
1100F — YAACGAGCGCAACCC
1492R — GGGTTACCTTGTTACGACTT
rambacV3F — CCTACGGGAGGCAGCAG
rambacV4R — GGACTACHVGGGTWTCTAAT
104F — GGCGVACGGGTGAGTAA
530R — CCGCNGCNGCTGGCAC
530F — GTGCCAGCMGCNGCGG
bac926R — CCGTCAATTYYTTTRAGTTT
1100R — GGGTTNCGNTCGTTR
18s rRNA Sequencing Primer List
EukV4F — CCAGCASCYGCGGTAATTCC
EukV4R — ACTTTCGTTCTTGATYRA
ionesV4R — ACTTTCGTTCTTGA
zigEukV4R — ACTTTCGTTCTTGATYRATGA
euk1391F — GTACACACCGCCCGTC
EukB-Rev — TGATCCTTCTGCAGGTTCACCTAC
Euk7F — AACCTGGTTGATCCTGCCAGT
Euk570R — GCTATTGGAGCTGGAATTAC
uni18sF — AGGGCAAKYCTGGTGCCAGC
uni18sR — GRCGGTATCTRATCGYCTT
nem18sF — CGATCAGATACCGCCCTAG
nem18sR — TACAAAGGGCAGGGACGTAAT
paraOxyF — GCYGCGGTAATWCCAGCTCT
paraoxyR — TGCNCTTCCGTCAATTYCTT
1080F — GGGRAACTTACCAGGTCC
1578R — GTGATRWGRTTTACTTRT
SSU316F — GCTTTCGWTGGTAGTGTATT
758R — CAACTGTCTCTATKAAYCG
AML1 — ATCAACTTTCGATGGTAGGATAGA
AML2 — GAACCCAAACACTTTGGTTTCC
wanda — CAGCCGCGGTAATTCCAGCT
960F — GGCTTAATTTGACTCAACRCG
1200R — GGGCATCACAGACCTG
1560F — TGGTGCATGGCCGTTCTTAGT
2035R — CATCTAAGGGCATCACAGACC
Euk60F — GAAACTGCGAATGGCTCATT
Euk515R — ACCAGACTTGCCCTCC
Euk516F — GGAGGGCAAGTCTGGT
Euk1055R — CGGCCATGCACCACC
eukss18F — CACCAGGTTGATTCTGCC
eukss530R — GTGCCAGCMGCCGCGG
Euk528F — CCGCGGTAATTCCAGCTC
EukR18R — CGTTATCGGAATTAACCAGAC
SSUF04 — GCTTGTAAAGATTAAGCC
SSUR22 — GCCTGCTGCCTTCCTTGGA
ITS Sequencing Primer List
ITS1F — CTTGGTCATTTAGAGGAAGTAA
ITS2R — GCTGCGTTCTTCATCGATGC
ITS4R — TCCTCCGCTTATTGATATGC
ITS3F — GCATCGATGAAGAACGCAGC
ITS3kyo2F — GATGAAGAACGYAGYRAA
ITS5F — GGAAGTAAAAGTCGTAACAAGG
ITS7 — GTGARTCATCGAATCTTTG
ITS9 — GAACGCAGCRAANNGYGA
ITS6pyth — GAAGGTGAAGTCGTAACAAGG
ITS7Rpyth — AGCGTTCTTCATCGATGTGC
gITS7F — GTGARTCATCGARTCTTTG
ITS4ngsR — TTCCTSCGCTTATTGATATGC
ramITS1ooF — CGGAAGGATCATTACCAC
ramITS58ooR — AGCCTAGACATCCACTGCTG
ENDONTSF — AAGGTCTCCGTAGGTGAAC
ENDONTSR — GTATCCCTACCTGATCCGAG
its58funFbar1 — AACTTTYRRCAAYGGATCWCT
its4funR — AGCCTCCGCTTATTGATATGCTTAART
symbiITS1 — GAATTGCAGAACTCCGTG
symbiITS2 — GGATCCATATGCTTAAGTTCAGCGGGT
traceITS1ooF — GGAAGGATCATTACCAC AC
Why Should I Choose the 16s Sequencing Service?
Prokaryotes today are divided into two domains, Archaea and Bacteria. These two domains are of particular interest in areas of research including:
The initial objective of the many studies within these fields among others is often the same; identify which microbes are present, or more importantly, which are absent. The 16s rRNA gene is an excellent sequencing target in order to complete such studies. There are nine hypervariable regions found in the 16s rRNA gene, and each of these regions is flanked by a highly conserved region. Our in-house 16s rRNA sequencing primer pairs are specifically designed to target these flanking conserved regions thereby allowing us at MR DNA to perform PCR amplification and DNA sequencing of your submitted microbial samples. Of the nine hypervariable regions found in the 16s rRNA gene, some regions may be better suited to complete certain phylogenetic studies over others. Feel free to consult our experts at MR DNA is order to determine which 16s rRNA primer pair is best suited to meet your sequencing needs.
The already cost effective method of 16s rRNA sequencing continues to reduce in cost as sequencing technology continues to advance. By utilizing the technology made available by next-generation sequencing platforms, we are able to generate the necessary data required to complete these 16s rRNA phylogenetic studies in a much more time efficient and cost-effective manner. For more information concerning our 16s rRNA sequencing capabilities, feel free to Contact us.
What is 16s rRNA Sequencing?
16s rRNA sequencing has become one of the leading methods for phylogenetic studies. The popularization of 16s sequencing methods has been due in large part to the wide availability of PCR and Next-generation sequencing facilities, such as MRDNA. But what is 16s rRNA sequencing? And why should you choose 16s sequencing methods over other DNA sequencing methods?
16s rRNA sequencing refers to sequencing the 16s rRNA gene that codes for the small subunit (SSU) of the ribosome found in prokaryotes such as Bacteria and Archaea. There are several factors that make the 16s rRNA gene the perfect target to complete your taxonomy or phylogeny studies.
Often times, researchers will have some confusion regarding the differences between 16s metagenomic sequencing methods and shotgun metagenomic sequencing methods. In short, shotgun metagenome sequencing is aptly named due to the fact that the goal of this DNA sequencing method is to sequence all genes from all organisms in a given sample. Whereas in the case of 16s metagenome sequencing, the goal is to sequence the 16s rRNA gene specifically.
Related Research
Periodontitis is caused by dysbiotic subgingival bacterial communities that may lead to increased bacterial invasion into gingival tissues. Although shifts in community structures associated with transition from health to periodontitis have been well characterized, the nature of bacteria present within the gingival tissue of periodontal lesions is not known. To characterize microbiota within tissues of periodontal lesions and compare them with plaque microbiota, gingival tissues and subgingival plaques were obtained from 7 patients with chronic periodontitis. A sequencing analysis of the 16S rRNA gene revealed that species richness and diversity were not significantly different between the 2 groups. However, intersubject variability of intratissue communities was smaller than that of plaque communities. In addition, when compared with the plaque communities, intratissue communities were characterized by decreased abundance of Firmicutes and increased abundance of Fusobacteria and Chloroflexi. In particular, Fusobacterium nucleatum and Porphyromonas gingivalis were highly enriched within the tissue, composing 15% to 40% of the total bacteria. Furthermore, biofilms, as visualized by alcian blue staining and atomic force microscopy, were observed within the tissue where the degradation of connective tissue fibers was prominent. In conclusion, very complex bacterial communities exist in the form of biofilms within the gingival tissue of periodontal lesions, which potentially serve as a reservoir for persistent infection. This novel finding may prompt new research on therapeutic strategies to treat periodontitis.
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