scholarly journals Species identification of clinically important Aeromonas spp. by restriction fragment length polymorphism of 16S rDNA

2007 ◽  
Vol 44 (5) ◽  
pp. 550-554 ◽  
Author(s):  
S. Ghatak ◽  
R.K. Agarwal ◽  
K.N. Bhilegaonkar
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Species Identification ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Restriction Fragment Length

Analysis of the Bacterial Diversity Existing on Animal Hide and Wool: Development of a Preliminary PCR-Restriction Fragment Length Polymorphism Fingerprint Database for Identifying Isolates

2012 ◽  
Vol 95 (6) ◽  
pp. 1750-1754 ◽  
Author(s):  
Chen Yu ◽  
Gao Hongwei ◽  
Zhang Yanming ◽  
Deng Mingjun ◽  
Wu Zhenxing ◽  
...  
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Nutrient Broth ◽  
Universal Primers ◽  
Comamonas Testosteroni ◽  
Restriction Fragment Length

Abstract Twenty-one bacterial strains were isolated from imported cattle hide and rabbit wool using two types of media, nutrient broth, and nutrient broth with serum. The bacteria identified were Brevibacillus laterosporus, Leclercia adecarboxylata, Peptococcus niger, Bacillus circulans, Raoultella ornithinolytica, Bacillus subtilis, Bacillus cereus, Bacillus thermobacillus, Bacillus choshinensis, Bacillus sphaericus, Acinetobacter haemolyticus, Sphingomonas paucimobilis, Bacillus thuringiensis, Staphylococcus intermedius, Mycobacteria, Moraxella, Klebsiella pneumoniae, Ralstonia pickettii, Staphylococcus chromogenes, Comamonas testosteroni, and Cupriavidus pauculus. The 16s rDNA gene of each bacterium was amplified using the universal primers 27f and 1492r. The amplicons were digested with AvaI, BamHI, BgII, DraI, EcoRI, EcoRV, HindIII, HinfI, HpaI, PstI, SmaI, TaqI, XbaI, XmaI, AluI, XhoI, and PvuI individually. A specific fingerprint from the PCR-restriction fragment length polymorphism method based on 16s rDNA was obtained for each bacterium. The results showed that the method developed was useful not only for bacterial identification but also for the etiological investigation of pathogens in imported animal hair and wool.

scholarly journals MOLECULAR IDENTIFICATION AND CLONAL RELATION OF ATYPICAL ISOLATE Aeromonas salmonicida USING RESTRICTION FRAGMENT LENGTH POLYMORPHISM (RFLP)

2011 ◽  
Vol 6 (2) ◽  
pp. 183
Author(s):  
Riza Priyatna ◽  
Indaryulianto Indaryulianto ◽  
Kurniasih Kurniasih
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Aeromonas Salmonicida ◽  
Freshwater Aquaculture ◽  
Restriction Sites ◽  
Restriction Fragment Length

Aeromonas salmonicida is responsible in many cases of furunculosis outbreaks resulting in economic loss of freshwater aquaculture. Four isolates of A. salmonicida have been isolated from fish collected in four different regions in Indonesia and its clonal relation has yet to be determined. In the research, A. salmonicida isolates and ATCC atypical isolate as the control had been checked for their clonal relation using Restriction Fragment Length Polymorphism (RFLP) method in which restriction enzyme of AluI, HaeIII, MboI, and EheI were used. PCR test results using the primers 16S rDNA amplicon gave a positive response to the 1300 bp band. The result of RFLP analysis showed that A. salmonicida atypical isolates from Indonesia represent subspecies smithia except isolates from C. macropomum in Yogyakarta in MS and 16S rDNA regions. Isolates from Jambi, Pontianak and Semarang showed a slight variation on enzyme restriction sites. Isolates number 2, 3, 4 and 5 had the same restriction sites using AluI enzyme with MS primer. The restriction enzymes that could give the best result for RFLP method of A. salmonicida were HaeIII, MboI and EheI.

scholarly journals Optimasi Isolasi Genom untuk Analisis Keragaman Mikrob pada Fermentasi Singkong "Peyem" dengan Teknik Terminal Restriction Fragment Length Polymorphism (T-RFLP)

2013 ◽  
Vol 18 (1) ◽  
Author(s):  
Tati Barus
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
16S Rdna ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Genomic Dna ◽  
Fragment Length Polymorphism ◽  
Dna Purification ◽  
Restriction Fragment Length

"Peyem" merupakan salah satu pangan fermentasi Indonesia. Kualitas pangan fermentasi bergantung pada aktivitas mikrob yang terdapat selama proses fermentasi berlangsung. Salah satu teknik molekuler yang telah banyak digunakan untuk menganalisis komunitas mikrob pada suatu habitat adalah teknik Terminal–Restriction Fragment Lenght Polymorphism (T-RFLP). Metode isolasi genom dan jenis primer yang digunakan pada saat PCR penting pada teknik T- RFLP dalam mengkaji komunitas mikrob. Oleh sebab itu, penelitian ini bertujuan untuk membandingkan empat metode isolasi genom dan membandingkan penggunaan dua set primer dalam mengkaji komunitas bakteri dari "Peyem" dengan teknik T-RFLP. Genom komunitas bakteri diisolasi dengan menggunakan empat metode, yaitu: 1) QIAamp DNA Stool Mini Kit (G1), 2) QIAamp DNA Stool Mini Kit + lisozim (G2), 3) Genomic DNA Purification Kit (G3), dan 4) Genomic DNA Purification Kit + lisozim (G4). Untuk mengamplifikasi 16S rDNA digunakan dua set primer, yaitu: 1) primer 27F-FAM dan 1492R, 2) primer 63F-FAM dan 1387R. Hasil penelitian menunjukkan isolasi genom dengan metode G4 menghasilkan konsentrasi genom tertinggi (330,20 ng/µl) dibandingkan metode G1, G2, dan G3 (163,50 ng/µl; 183,25 ng/µl, dan 260,80 ng/µl). Primer 27F-FAM menghasilkan jumlah peak yang lebih tertinggi (264) dibandingkan dengan primer 63F-FAM (177). Jumlah peak TRF pada teknik TRFLP menggambarkan keragaman komunitas mikrob. Dengan demikian isolasi genom dengan Genomic DNA Purification Kit + lysozyme dan penggunaan pasangan primer 27F-FAM-1492R adalah yang terbaik untuk menganalisis komunitas bakteri dari "Peyem" dengan teknik T-RFLP.Kata kunci: Genom, Primer, T-RFLP, Mikrob, "Peyem"

Sign in / Sign up

Export Citation Format

Share Document