TLR2KO Morphine 2
Mus musculus domesticus
Protocols: Fecal content was collected from gut region encompassing distal cecum and approximately one inch of the colon and frozen on dry ice. The fecal matter was lysed using glass beads in MagnaLyser tissue disruptor (Roche) and total DNA isolated using Power-soil/fecal DNA isolation kit (Mo-Bio) as per manufacturers specifications. All samples was quantified via the Qubit Quant-iT dsDNA Broad- Range Kit (Invitrogen, Life Technologies, Grand Island, NY) to ensure that they met minimum concentration and mass of DNA and submitted to either Second genome Inc. or University of Minnesota Genomic Center for microbiome analysis. Slow release morphine pellets (25mg; ~1M serum levels of morphine for 5-6 days) and corresponding placebo or naltrexone pellets, as appropriate, were kindly provided by National Institute of Drug Abuse (NIDA, National Institutes of Health, Rockville, MD). The implantation procedure involved 3% isoflurane induced anesthesia, followed by making a small incision at the dorsal torso of the mice. The appropriate pellet was inserted into the small pocket created during incision and the wound was closed using stainless steel wound-clips. The whole process was carried out under aseptic conditions. To enrich the sample for the bacterial 16S V4 rDNA region, DNA was amplified utilizing fusion primers designed against the surrounding conserved regions which are tailed with sequences to incorporate Illumina (San Diego, CA) flow cell adapters and indexing barcodes. Each sample was PCR amplified with two differently bar coded V4 fusion primers and were advanced for pooling and sequencing. For each sample, amplified products were concentrated using a solid-phase reversible immobilization method for the purification of PCR products and quantified by electrophoresis using an Agilent 2100 Bioanalyzer. Using QIIME, sequences were quality filtered and demultiplexed using exact matches to the supplied DNA barcodes.
ENA first public
2016-06-01
ENA last update
2015-12-10
External Id
SAMEA3706985
INSDC center alias
Department of Surgery, BTR University of Minnesota
INSDC center name
Department of Surgery, BTR University of Minnesota
INSDC first public
2016-06-01T17:01:04Z
INSDC last update
2015-12-10T16:33:54Z
INSDC status
public
Submitter Id
E-MTAB-37222:TLR2KO Morphine 2
broker name
ArrayExpress
common name
western European house mouse
organism part
feces
sample name
E-MTAB-37222:TLR2KO Morphine 2