Microbiome Core Facility

Bioinformatics analysis of 16S rRNA amplicon sequencing data

Illumina sequencing options:
MiSeq SE5(50) $1,491
MiSeq PE150(300) $2,020
MiSeq PE250(500) $2,315
MiSeq PE300(600) $2,891

HiSeq High output SE50 $1,709
HiSeq High output PE50 $2,891
HiSeq High output SE100 $2,720
HiSeq High output PE100 $3,575
HiSeq High output PE150 $3,668

HiSeq RAPID SE50 $1,861
HiSeq RAPID PE50 $3,186
HiSeq RAPID SE100 $2,720
HiSeq RAPID PE100 $3,808
HiSeq RAPID PE150 $4,461
HiSeq RAPID PE250 $5,130

HiSeq 4000 SE50 $2,328
HiSeq 4000 PE50 $2,953
HiSeq 4000 PE100 $3,497
HiSeq 4000 SE100 $2,955
HiSeq 4000 SE150 $3,577
HiSeq 4000 PE150 $3,808
* prices are for UNC, NC State or Federal Funded / all other

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.

RNA sequencing (see Illumina sequencing for service options)
Library preparation: $364
RNA extraction: $28

Clone library creation:
In order to confirm bacterial identity, PCR-amplified 16S rRNA gene 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