Reference - Detail
| RRC ID | 21633 |
|---|---|
| Author | Crosbie ND, Pöckl M, Weisse T. |
| Title | Rapid establishment of clonal isolates of freshwater autotrophic picoplankton by single-cell and single-colony sorting. |
| Journal | J Microbiol Methods |
| Abstract |
We describe single-cell and single-colony sorting protocols which allowed for rapid establishment of a diverse culture collection of clonal autotrophic picoplankton (APP) isolates originating from oligotrophic and oligo-mesotrophic subalpine lakes. Overall sort recoveries, expressed as the percentage of sorted microwells exhibiting APP growth, ranged from 5% to 17% depending on the type of APP, but the growth success varied greatly (from 0% to 68%) depending on the origin of the sorted sample. We applied two direct sequencing and two denaturing gradient gel electrophoresis (DGGE) protocols to identify and characterize the genetic purity of 21 of our picocyanobacteria cultures, namely, direct sequencing of the 16S rRNA gene and cpcBA-IGS region, and DGGE analyses involving a 194-bp fragment of the internal transcribed spacer (ITS) and a ca. 500-bp fragment of the phycocyanin (PC) operon (cpcBA-IGS, novel protocol described herein). Of those 21 picocyanobacteria cultures obtained by single-cell/single-colony sorting and subsequently characterized genetically/screened for genetic purity, only one culture was composed of multiple picocyanobacterial strains. |
| Volume | 55(2) |
| Pages | 361-70 |
| Published | 2003-11-1 |
| DOI | 10.1016/s0167-7012(03)00167-2 |
| PII | S0167701203001672 |
| PMID | 14529957 |
| MeSH | Animals Base Sequence DNA / chemistry DNA / genetics DNA, Ribosomal Spacer / chemistry DNA, Ribosomal Spacer / genetics Electrophoresis Flow Cytometry Fresh Water Microscopy, Fluorescence Molecular Sequence Data Phycocyanin / chemistry Phycocyanin / genetics Plankton / classification Plankton / genetics Plankton / growth & development* RNA, Ribosomal, 16S / chemistry RNA, Ribosomal, 16S / genetics Sequence Analysis, DNA Water Microbiology* |
| IF | 1.707 |
| Times Cited | 38 |
| WOS Category | BIOCHEMICAL RESEARCH METHODS MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| Total number of mentions | 0 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Algae | NIES-1345 NIES-1344 NIES-1343 NIES-1341 NIES-1342 NIES-1346 NIES-1347 NIES-1348 |