Lab Members:
-
M. Andrea Azcarate Peril
ASSISTANT PROFESSOR
Microbiome Core Facility
- 11 Services
- 1 Member
The human body is the home to approximately 10,000,000,000 bacterial cells that inhabit our gastrointestinal tract, mouth, urogenital tract, and our skin. Most of these bacteria are uncultivable, which meant in the past that researchers were unable to study and characterize them. The development of metagenomic technologies allow us today to examine microbial communities through culture independent techniques, of which the most used are next-generation sequencing methods.
The UNC Microbiome Core Facility was established to provide technical and research support to study community structure and function of host-associated microbial populations.
Bioinformatics analysis of 16S amplicon sequencing data
UNC/NC State or covered by federal funds $64
non UNC/NC State or not covered by federal funds $68
Bioinformatics analysis of 16S amplicon sequencing data
UNC/NC State or covered by federal funds $64
non UNC/NC State or not covered by federal funds $68
Isolation of total DNA.
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
Isolation of total DNA.
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
Illumina sequencing
MiSeq SE5(50) $823 $1,193
MiSeq PE150(300) $1023 $1,483
MiSeq PE250(500) $1125 $1,631
MiSeq PE300(600) $1478 $2,143
HiSeq High output SE50 $913 $1,324
HiSeq High output PE50 $1503 $2,179
HiSeq High output SE100 $1238 $1,795
HiSeq High output... Show more »
Illumina sequencing
MiSeq SE5(50) $823 $1,193
MiSeq PE150(300) $1023 $1,483
MiSeq PE250(500) $1125 $1,631
MiSeq PE300(600) $1478 $2,143
HiSeq High output SE50 $913 $1,324
HiSeq High output PE50 $1503 $2,179
HiSeq High output SE100 $1238 $1,795
HiSeq High output PE100 $2153 $3,122
HiSeq RAPIDSE50 $1073 $1,556
HiSeq RAPIDPE50 $1827 $2,649
HiSeq RAPIDSE100 $1561 $2,263
HiSeq RAPIDPE100 $2802 $4,063
HiSeq RAPIDPE150 $3265 $4,734
The Ion Torrent sequencing platform from Life Technologies employs First PostLight™ sequencing technology (semiconductor technology). The detection of a specific nucleotide on a growing DNA strand occurs inside a well of an Ion Torrent semiconductor chip. The incorporation of the correct base triggers a chemical reaction that releases... Show more »
The Ion Torrent sequencing platform from Life Technologies employs First PostLight™ sequencing technology (semiconductor technology). The detection of a specific nucleotide on a growing DNA strand occurs inside a well of an Ion Torrent semiconductor chip. The incorporation of the correct base triggers a chemical reaction that releases a hydrogen ion as byproduct, which in turn changes the pH of the solution. The Ion sensor captures voltage change measurements from the direct release of hydrogen ions following DNA polymerization and incorporation events get recorded as digital signals 1 and 0. This technology differs from other sequencing technologies in that no modified nucleotides, no lasers, no optics, no camera, no fluorescence, no enzyme cascade are used.
Ion Torrent sequencing fees:
314 (400bp read)
UNC/NC State/ Covered by federal funds $767
Non-UNC or NC State or not covered by federal funds $1,212
316 (400 bp read)
UNC/NC State/ Covered by federal funds $1,057
Non-UNC or NC State or not covered by federal funds $1,607
318 (400 bp read)
UNC/NC State/ Covered by federal funds $1,464
Non-UNC or NC State or not covered by federal funds $2,225
Clone library creation:
In order to confirm bacterial identity, PCR-amplified 16S rDNA fragments can be cloned into plasmids and transformed into an appropriate host (usually E. coli). Each transformed E. coli colony will contain only one 16S rDNA PCR-originated fragment from the original sample. A number of colonies are picked... Show more »
Clone library creation:
In order to confirm bacterial identity, PCR-amplified 16S rDNA fragments can be cloned into plasmids and transformed into an appropriate host (usually E. coli). Each transformed E. coli colony will contain only one 16S rDNA PCR-originated fragment from the original sample. A number of colonies are picked randomly and the insert is sequenced. The DNA sequence is then compared to a database (for example RDP II) for identification by DNA homology.
Equipment:
Genetix QPix2 XT Colony Picker System
UNC/NC State/ Covered by federal funds $480
Non-UNC or NC State or not covered by federal funds $528
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
UNC/NC State/ Covered by federal funds $16
Non-UNC or NC State or not covered by federal funds $25
- Illumina RNA Library preparation with barcoding
UNC/NC State/ Covered by federal funds $220
Non-UNC or NC State or not covered by federal funds $319
- Illumina RNA Library preparation with barcoding
UNC/NC State/ Covered by federal funds $220
Non-UNC or NC State or not covered by federal funds $319
Pyrosequencing is a DNA sequencing technique that utilizes enzyme coupled reactions and bioluminescence to monitor the pyrophosphate release accompanying nucleotide incorporation, in real time. The pyrosequencing method was developed by the group of Pal Nyren. With the aim of commercializing the new DNA sequencing technique, Nyren and... Show more »
Pyrosequencing is a DNA sequencing technique that utilizes enzyme coupled reactions and bioluminescence to monitor the pyrophosphate release accompanying nucleotide incorporation, in real time. The pyrosequencing method was developed by the group of Pal Nyren. With the aim of commercializing the new DNA sequencing technique, Nyren and others founded Pyrosequencing AB in 1997. The technology was licensed to 454 Life Sciences, which was later acquired by Roche in 2007. 16S DNA amplicon sequencing enables the identification and quantification of bacteria within complex mixtures of PCR amplicons.
454 amplicon pyrosequencing fees:
Full plate
UNC/NC State/ Covered by federal funds $9,797
Non-UNC or NC State or not covered by federal funds $10,758
TRFLP is a method that allows the identification of major phylogenetic groups in complex microbial communities. It is based on the treatment with restriction enzymes of a DNA fragment specifically amplified by PCR. The terminal fluorescent-labeled fragments produced are separated by size using a DNA sequencer. Each specific bacterial... Show more »
TRFLP is a method that allows the identification of major phylogenetic groups in complex microbial communities. It is based on the treatment with restriction enzymes of a DNA fragment specifically amplified by PCR. The terminal fluorescent-labeled fragments produced are separated by size using a DNA sequencer. Each specific bacterial group will produce a specific profile that can be identified by comparing them to existing profile databases.
UNC/NC State/ Covered by federal funds $130
Non-UNC or NC State or not covered by federal funds $150
The method is based on the conserved 16S rDNA gene present in all bacteria (a comprehensive list of 16S rDNA sequences from gut bacteria have been deposited and categorized in the Ribosomal Database Project II ). By calculating the number of conserved 16S rDNA genes, qPCR can indirectly indicate the number of bacteria present in a... Show more »
The method is based on the conserved 16S rDNA gene present in all bacteria (a comprehensive list of 16S rDNA sequences from gut bacteria have been deposited and categorized in the Ribosomal Database Project II ). By calculating the number of conserved 16S rDNA genes, qPCR can indirectly indicate the number of bacteria present in a given sample. Additionally, appropriate design of qPCR oligonucleotide probes (primers) that can identify the sequence of the 16S gene variable regions allows for the amplification of genes from a specific bacterium of interest whilst excluding all other bacteria, making qPCR a useful tool for quantifying specific bacteria in a complex microbial community.
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Fees:
- qPCR one-time set up fee
UNC/NC State/ Covered by federal funds $118
Non-UNC or NC State or not covered by federal funds $169
- qPCR
UNC/NC State/ Covered by federal funds $21
Non-UNC or NC State or not covered by federal funds $28
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