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UncleRimas IP-: frRJnGOEbQQ6o |
DNA from Lactobacillus isolates was extracted and was slot-blotted onto Nytran nylon membranes and probed with whole genomic DNA probes prepared from known American Type Culture Collection (ATCC) strains. , , ( -?) , . By randomly labelling the whole genomic DNA with [ -32P]dATP using the Random Primed DNA Labelling Kit, whole chromosomal probes were made from ATCC Lactobacillus strains. ( ?) Lactobacillus, / ATCC, [α-32P]dATP P.S. , . |
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-[α-32P]dATP- . -...... |
UncleRimas IP-: frRJnGOEbQQ6o |
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guest: IP-: frwHRv5zEzGDY |
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UncleRimas IP-: frRJnGOEbQQ6o |
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Pharmaol |
[-32P]dATP , Random Primed DNA Labelling Kit. |
guest: IP-: frwHRv5zEzGDY |
. . , , ( ) . , , , . |
UncleRimas IP-: frRJnGOEbQQ6o |
, . Each sample was evently aliquoted into 3 adjacent wells and was vacuum-transferred to the membrane (Nytran nylon membranes). , 3- ? ! - , ) |
Pharmaol |
, (). (UncleRimas @ 14.04.2013 22:01) Each sample was evently aliquoted into 3 adjacent wells and was vacuum-transferred to the membrane (Nytran nylon membranes). , 3- ? " " 3 . |
UncleRimas IP-: frRJnGOEbQQ6o |
? , , . 5% , 48 6% CO2 37C. , 0.5% [/] , 1% [/] , 0.02% -80, 1% [/ l] , 1% [/] [7]. 20 2000 , TES (30 , 5 , 50 NaCl [pH 8.0]), 1 (25% , 50 T, 1 [pH 8.0]) 60 37 : 35 (Sigma) 30 , 16 ( XIV , Streptomyces griseus) (Sigma) 1 , 1 0.25 M (pH 8.0) 5 25. 0.4 20% 60 65C . 10 (Sigma), 30 . - . 95% - 22C. 70% , , 10 -Cl 1 (pH 8.0). |
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UncleRimas IP-: frRJnGOEbQQ6o |
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UncleRimas IP-: frRJnGOEbQQ6o |
, , RAPD- RAPD- - ? , " , RAPD-", ? The RAPD types isolated from blood agar (subjects I, O, T, and U) showed no similarity with those of any of the isolates from Rogosa agar but had high similarity with the RAPD pattern from the type strain of L. iners (Table 1). No single subject was dominated by more than two RAPD types. Some of the RAPD types were recurrent; in addition, two subjects (G and H) harbored the same species but with different RAPD types (Table 1). . "harbored" - , )) ? RAPD.. () , ! ! |
guest: IP-: fr/9gxL6jBvmM |
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UncleRimas IP-: frRJnGOEbQQ6o |
TTGE- : "TTGE pattern showed a slight deviation in the position of bands" - (?) ? PCR-G was used to group the lactobacilli with primers Ldel-7, Lac-2 etc.. PCR II-1 with the primers etc... PCR II-2 with primers etc.. . , PCR-G , , G-. PCR II-1, PCR II-2 ? . ) |
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"PCR-G" - G-attachment protein. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC263972/pdf/jcm00009-0201.pdf PCR II-1, PCR II-2 - II. http://libgen.org/scimag2/10.1111/j.1574-6968.2000.tb09155.x.pdf
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UncleRimas IP-: frRJnGOEbQQ6o |
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UncleRimas IP-: frRJnGOEbQQ6o |
The vaginal flora of most participants in the study was dominated by a single RAPD type, but five of them harbored two RAPD types representing two different species or strains. : , RAPD , , , RAPD , . |
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UncleRimas IP-: frRJnGOEbQQ6o |
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UncleRimas IP-: frRJnGOEbQQ6o |
With this objective, we applied PCR-based methods for the identification of the Lactobacillus flora. Randomly amplified polymorphic DNA (RAPD) analysis (10, 22) was used to group the isolates; temporal temperature gradient gel electrophoresis (TTGE), where the 16S ribosomal DNA (rDNA) molecule is used, was used for identification to the species level (29); and multiplex PCR, where another area of the rDNA, namely, the intergenic spacer region between the 16S and the 23S rDNAs, is used for the identification (27), was also used. : - . (RAPD ) (10, 22); - (TTGE) 16S () (29); , , 16S-23S (27). |
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