Amino-acid analysis was performed with a ninhydrin-based SYKAM System S4300 Amino Acid Analyzer (SYKAM, Germany). Molecular cloning. Total RNA was extracted from dissected mussel feet regions that tested positive in the Arnow assay using an RNeasy Mini Kit (QIAGEN, Valencia, CA, USA). Briefly...
RNA extraction, cDNA synthesis, and realtime-qPCR Total RNA was extracted from the tissue samples accord- ing to the manufacturer's instructions using the RNeasy plus Universal kit (Qiagen, Hilden, Germany). The con- centration and purity of the isolated RNA were assessed using a NanoDrop ...
Nucleotide sequences from fragments of four parts of the protein encoding gene, CAD (=Rudimentary in Drosophila melanogaster), and from the mitochondrial cytochrome oxidase I gene were obtained from each specimen and extracted using the DNeasy DNA extraction kit (QIAGEN, Inc.). The standard protoco...
Ni-TA resin (Qiagen) was preloaded with importin-α in binding buffer for 1 h at 4°C and washed three times with binding buffer. Crude extracts were generated from digitonin-treated control or stressed cells in binding buffer containing 0.5 mM DTT, 0.5% SDS and protease inhibitors (...
The plasmid pCR2.1TOPOgdhA is cleaved with the enzyme EcoRI, the cleavage batch is separated on 0.8% agarose gel and the gdhA fragment 2.1 kbp in size is isolated with the aid of the “QIAquick Gel Extraction Kit” (QIAGEN, Hilden, Germany). The plasmid pMW218 (Nippon Gene, Toyama, Ja...
The PCR amplicons from conventional PCRs for Bartonella and Rickettsia were purified using a QIAquick PCR purification kit (Qiagen, Valencia, CA) according to the manufacturer’s instructions and then sequenced in both directions by using an ABI 3130 genetic analyzer (Applied Biosystems, Foster City...
the background prevalence is typically less than 5% and a prevalence over 40% may suggest an outbreak [33]; however, we note that these recommendations are mostly based on amphibian studies because surveys of ranavirus in wild reptiles are limited. In addition to having a low detection rate ...
PCR products were subsequently purified using the QIAquick® PCR Purification Kit (Qiagen, Mississauga, ON, Canada) and inserted into the pCR®II-TOPO® (ThermoFisher Scientific; Waltham, MA, USA) vector, as recommended by the supplier. These Ugt2b cDNA were sub-cloned into the NheI/...