The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2003
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | München, Techn. Univ., Diss., 2003 |
| Beschreibung: | VII, 128 S. Ill., graph. Darst. |
Internformat
MARC
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| 100 | 1 | |a Zwirglmaier, Katrin |d 1974- |e Verfasser |0 (DE-588)128512717 |4 aut | |
| 245 | 1 | 0 | |a The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms |c Katrin Zwirglmaier |
| 264 | 1 | |c 2003 | |
| 300 | |a VII, 128 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Techn. Univ., Diss., 2003 | ||
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Datensatz im Suchindex
| _version_ | 1807682894714372096 |
|---|---|
| adam_text |
TABLE
OF
CONTENTS
A.
INTRODUCTION
1
B.
MATERIAL
AND
METHODS
B.L.
ORGANISMS
5
B.2.
CULTIVATION
OF
ORGANISMS
7
B.3.
PRIMERS
8
B.4.
PROBES
10
B
.5.
ISOLATION
OF
NUCLEIC
ACIDS
12
B.5.1
.
ISOLATION
OF
GENOMIC
DNA
12
B.5.2.
ISOLATION
OF
PLASMID
DNA
13
B.6.
PHOTOMETRIC
MEASUREMENT
OF
NUCLEIC
ACID
SOLUTIONS
14
B.7.
AGAROSE
GEL
ELECTROPHORESIS
15
B.8.
POLYMERASE
CHAIN
REACTION
(PCR)
16
B.9.
LABELLING
OF
NUCLEIC
ACIDS
WITH
BIOTIN,
DIGOXIGENIN
OR
FLUORESCENT
DYES
18
B.9.
1.
OLIGONUCLEOTIDE
PROBES
18
B.9.2.
POLYNUCLEOTIDE
RNA
PROBES
-
IN
VITRO
TRANSCRIPTION
18
B.10.
CELL
FIXATION
20
B.
10.1.
CELL
FIXATION
WITH
PFA
20
B.10.2.
CELL
FIXATION
WITH
ETHANOL
20
B.I0.3.
CELL
FIXATION
WITH
FORMALDEHYDE
/
ACETIC
ACID
21
B.10.4.
CELL
FIXATION
WITH
METHANOL
21
B.
10.5.
POSTFIXATIONAL
TREATMENTS
22
B.
10.5.1.
HEAT
FIXATION
22
B.
10.5.2.
LYSOZYME
22
B.
10.5.3.
PROTEINASE
K
23
B.L
1.
HYBRIDISATION
TECHNIQUES
23
B.
11.1.
EVALUATION
OF
HYBRIDISATION
CONDITIONS
23
B
.
11.2.
HYBRIDISATION
WITH
OLIGONUCLEOTIDE
PROBES
25
B.11.3.
HYBRIDISATION
WITH
RNA
POLYNUCLEOTIDE
PROBES
26
B
.
11.3.1.
PROBES
TARGETING
RRNA
26
B.L
1.3.2.
PROBES
TARGETING
PLASMIDS
27
B.L
1.3.3.
PROBES
TARGETING
GENOMIC
DNA
27
B.12.
CELL
SORTING
28
B.12.1.
MAGNETIC
CELL
SORTING
(MACS)
28
B.
12.2.
CELL
SORTING
IN
MICROPLATES
28
B.
12.2.1.
STREPTAVIDIN
COATED
MICROPLATES
28
B.
12.2.2.
DNA
COATED
MICROPLATES
29
B.
12.2.2.1.
COATING
OF
MICROPLATES
WITH
DNA
29
B.
12.2.2.2.
CELL
SORTING
(HYBRIDISATION
IN
COATED
MICROPLATES
-
HYCOMP)
30
B.13.
DETECTION
OF
CELLS
AFTER
HYBRIDISATION
31
B.13.1.
DETECTION
IN
MICROPLATES
31
B.13.2.
DETECTION
FOR
MICROSCOPE
ANALYSIS
32
B.14.
QUANTIFICATION
OF
CELL
SORTING
EFFICIENCY
32
B.14.1.
SEMIQUANTITATIVE
PCR
32
B.
14.2.
MANUAL
CELL
COUNTS
33
B.
14.3.
COMPUTER
BASED
QUANTIFICATION
34
B.15.
SOFTWARE
34
C.
RESULTS
PART
I
-
FLUORESCENCE
IN
SITU
HYBRIDISATION
(FISH)
WITH
POLYNUCLEOTIDE
TRANSCRIPT
PROBES
C.
1.
GROUPS
OF
ORGANISMS
TARGETED
WITH
POLYNUCLEOTIDE
PROBES
36
C.
1.1.
GRAM-NEGATIVE
BACTERIA
38
C.
1.2.
GRAM-POSITIVE
BACTERIA
40
C.
1.3.
YEASTS
44
C.
1.4.
MAMMALIAN
CELLS
50
C.2.
CLASSES
OF
TARGET
MOLECULES
FOR
POLYNUCLEOTIDE
PROBES
52
C.2.1
.
POLYNUCLEOTIDE
PROBES
TARGETING
RRNA
52
C.2.
1.1.
STANDARD
POLYNUCLEOTIDE
PROBES
52
C.2.
1.2.
EVALUATION
OF
PROBE
LENGTH
52
C.2.1
.3.
INFLUENCE
OF
SECONDARY
STRUCTURE
ON
HYBRIDISATION
SIGNAL
54
C.2.2.
POLYNUCLEOTIDE
PROBES
TARGETING
PLASMIDS
57
C.2.3.
POLYNUCLEOTIDE
PROBES
TARGETING
CHROMOSOMAL
DNA
65
C.2.3.1.
GLYCEROL
ALDEHYDE
3-PHOSPHATE
DEHYDROGENASE
(GAPDH)
65
C.2.3.2.
XANTHOMONAS
CAMPESTRIS
PATHOVARS
68
PART
II
-
CELL
SORTING
WITH POLYNUCLEOTIDE
PROBES
C.3.
CELL
SORTING
METHODS
70
C.4.
OPTIMISATION
OF
THE
REACTION
PARAMETERS
72
C.4.1.
SAMPLE
PREPARATION
72
C.4.2.
PARAMETERS
FOR
1
ST
HYBRIDISATION
73
C.4.2.1.
STRINGENCY
73
C.4.2.2.
HYBRIDISATION
TIME
74
C.4.2.3.
AMOUNT
OF
PROBE
74
C
.4.3.
PARAMETERS
FOR
2
ND
HYBRIDISATION
76
C.4.3.1.
STRINGENCY
76
C.4.3.2.
HYBRIDISATION
TIME
76
C.5.
CELL
SORTING
EFFICIENCY
77
C.5.
1.
EVALUATION
VIA
SEMIQUANTITATIVE
PCR
77
C.5.2.
QUANTIFICATION
VIA
MANUAL
CELL
COUNTS
78
C.
5.3.
COMPUTER
BASED
QUANTIFICATION
78
C.6.
APPLICATING
THE
METHOD
TO
ENVIRONMENTAL
SAMPLES
80
D.DISCUSSION
PART
I
-
FLUORESCENCE
IN
SITU
HYBRIDISATION
(FISH)
WITH
POLYNUCLEOTIDE
TRANSCRIPT
PROBES
D.
1.
TARGET
ORGANISMS
83
D.2.
TARGET
MOLECULES
85
D2.1.
RRNA
85
D.2.2.
DNA
:
PLASMIDS
AND
CHROMOSOMAL
DNA
-
RING-FISH
86
D.3.
NETWORK
HYPOTHESIS
93
D.4.
PROBE
DESIGN
99
D.5.
POSSIBLE
APPLICATIONS
100
PART
N
-
CELL
SORTING
WITH
POLYNUCLEOTIDE
PROBES
D.6.
COMPARISON
OF
CELL
SORTING
METHODS
101
D.7.
CELL
SORTING
IN
DNA
COATED
MICROPLATES
102
D
.7.1.
CELL
FIXATION
.
102
D.7
.2.
HYBRIDISATION
PARAMETERS
102
D.7.3.
CELL
SORTING
EFFICIENCY
103
D.8.
ENVIRONMENTAL
SAMPLES
103
D.9.
POSSIBLE
APPLICATIONS
104
E.
SUMMARY
SUMMARY
(ENGLISH)
105
ZUSAMMENFASSUNG
(GERMAN)
106
F,
APPENDIX
107
G.
REFERENCES
118 |
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| discipline | Biologie |
| format | Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV017463433 |
| illustrated | Illustrated |
| indexdate | 2024-08-18T00:25:47Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-010521830 |
| oclc_num | 645502903 |
| open_access_boolean | |
| owner | DE-91 DE-BY-TUM DE-M49 DE-BY-TUM DE-12 |
| owner_facet | DE-91 DE-BY-TUM DE-M49 DE-BY-TUM DE-12 |
| physical | VII, 128 S. Ill., graph. Darst. |
| publishDate | 2003 |
| publishDateSearch | 2003 |
| publishDateSort | 2003 |
| record_format | marc |
| spelling | Zwirglmaier, Katrin 1974- Verfasser (DE-588)128512717 aut The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms Katrin Zwirglmaier 2003 VII, 128 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2003 Polynucleotide (DE-588)4268231-9 gnd rswk-swf Mikroorganismus (DE-588)4039226-0 gnd rswk-swf Fluoreszenz-in-situ-Hybridisierung (DE-588)4528641-3 gnd rswk-swf Sonde (DE-588)4055510-0 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Mikroorganismus (DE-588)4039226-0 s Sonde (DE-588)4055510-0 s Polynucleotide (DE-588)4268231-9 s Fluoreszenz-in-situ-Hybridisierung (DE-588)4528641-3 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010521830&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zwirglmaier, Katrin 1974- The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms Polynucleotide (DE-588)4268231-9 gnd Mikroorganismus (DE-588)4039226-0 gnd Fluoreszenz-in-situ-Hybridisierung (DE-588)4528641-3 gnd Sonde (DE-588)4055510-0 gnd |
| subject_GND | (DE-588)4268231-9 (DE-588)4039226-0 (DE-588)4528641-3 (DE-588)4055510-0 (DE-588)4113937-9 |
| title | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms |
| title_auth | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms |
| title_exact_search | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms |
| title_full | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms Katrin Zwirglmaier |
| title_fullStr | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms Katrin Zwirglmaier |
| title_full_unstemmed | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms Katrin Zwirglmaier |
| title_short | The use of polynucleotide RNA probes for detection, identification and cell sorting of microorganisms |
| title_sort | the use of polynucleotide rna probes for detection identification and cell sorting of microorganisms |
| topic | Polynucleotide (DE-588)4268231-9 gnd Mikroorganismus (DE-588)4039226-0 gnd Fluoreszenz-in-situ-Hybridisierung (DE-588)4528641-3 gnd Sonde (DE-588)4055510-0 gnd |
| topic_facet | Polynucleotide Mikroorganismus Fluoreszenz-in-situ-Hybridisierung Sonde Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010521830&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT zwirglmaierkatrin theuseofpolynucleotidernaprobesfordetectionidentificationandcellsortingofmicroorganisms |