| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The PCR was carried out in 50 μL reaction containing 40 ng of DNA, 0.2 μM of primers D602F and D753Rext (Integrated DNA Technology, USA), 0.2 mM dNTPs (Roche Diagnostics, New Zealand), 1× PCR reaction buffer, 2 U of Taq polymerase (Roche Diagnostics, New Zealand), 0.24 μg of BSA (Sigma, New Zealand) and 2.5 mM MgCl2. PCRs were performed in a PTC-200 Peltier Thermal Cycler (MJ Research Inc., USA) under the following cycling conditions; 94 °C for 2 min followed by 40 cycles of 94 °C for 20 s, 55 °C for 60 s and 72 °C for 2 min, with a final extension of 72 °C for 5 min. Amplicons of the correct size were purified using a GenScript PCR purification kit (GenScript, USA) and sequenced bi-directionally using a Applied Biosystems 3130xl Genetic Analyzer and the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, USA). | Get A Quote |
Didymosphenia geminata is a large, invasive, freshwater diatom that can produce distinctive and robust mucilaginous stalks. Over the last two decades, there has been a worldwide increase in the distribution and severity of D. geminata blooms. These dense, persistent blooms can have severe impacts on native species and ecosystem functioning. D. geminata is usually identified by microscopic methods that are time consuming, resource intensive, and dependent upon expert taxonomic identification, so the extent of surveillance programs has been limited. As an alternative, we have developed a TaqMan quantitative polymerase chain reaction(QPCR) assay for sensitive and rapid detection and enumeration of D. gem... More
