Nanodrop bioanalyzer nd1000

Before starting the present experiment, the shoots of transgenic olive lines were tested again for kanamycin resistance, at 150 mg L⁻¹ kanamycin-enriched media, and the presence of the transgenic genes in their DNA were ascertain. Olive genomic DNA was extracted from young leaves (150m g of fresh tissue) using a based CTAB (cetyl-trimethyl-ammonium bromide) method [47 ]. DNA concentration and quality were determined by 1% agarose gel electrophoresis and using a Nanodrop Bioanalyzer ND1000 (Thermo Scientific). Then, the transformant was screened for the transgene by polymerase chain reaction (PCR) using the osmotin-specific primers Osm-F (5′-CCAACAACCCAACTTGTTAAAA-3′) and Osm-R (5′-CGACAGAATAATTTGACCAAAAG-3′).
The PCR conditions were 95 °C for 5 min; 35 cycles of 94 °C for 30 s, 60 °C for 45 s, and 72 °C for 80 s, and final extension at 72 °C for 10 min. The reaction products were separated electrophoretically on 1.5% (w/v) agarose gel, stained with ethidium bromide, and photographed with a digital camera (Nikon coolpix 5700) (Figure 7).

Genomic DNA was isolated from the propagated phages according to the procedure described by Kieser et al. [28 ]. DNA quality was assessed using a Nanodrop Bioanalyzer ND1000 (ThermoScientific). Sequencing libraries were prepared by shearing 1 μg of DNA in blunt-ended fragments by linking the Ion adapters using an Ion XpressTM Plus Fragment Library Kit (Life Technologies, Carlsbad, USA) according to the manufacturer’s specifications. The sized and ligated fragments were amplified by emulsion-PCR using the Ion OneTouch 200 Template kit (Life Technologies, Carlsbad, USA). Quality and insert size distribution were assessed using an Agilent Bioanalyzer DNA 1000 chip. Libraries were sequenced on an Ion Torrent PGM semiconductor sequencer (Life Technologies, Carlsbad, USA) using the 200 bp protocol and an Ion Torrent 314 chip following the manufacturer instructions (Life Technologies, Carlsbad, USA).