| 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
|
| Resource |
| Algae |
NIES-1345
NIES-1344
NIES-1343
NIES-1341
NIES-1342
NIES-1346
NIES-1347
NIES-1348 |
