China
Africa
Explore chapters and articles related to this topic
Beneficial Lactic Acid Bacteria
Published in K. Balamurugan, U. Prithika, Pocket Guide to Bacterial Infections, 2019
Restriction fragment length polymorphism (RFLP) is characterized by the use of restriction enzymes to digest DNA and following separation of the restriction fragments according to their length by agarose gel electrophoresis (Adzitey et al. 2013). Single digestion with AciI of rpoB gene (coding for RNA polymerase β-subunit) in Leuconostoc, Oenococcus, Pediococcus, and two or three digestions (AciI, HinfI and MseI) in Lactobacillus spp. allowed to identify LAB species commonly isolated from wine (Claisse et al. 2007). Restriction patterns of the tuf gene, encoding the elongation factor Tu and universally distributed in gram-positive bacteria, derived by enzymes AluI and HaeIII could effectively differentiate closely related Lactobacillus species (Park et al. 2012). However, sometimes strain variations could not be demonstrated by the RFLP analysis. Morphologic differences (colony shape and size) were evident between Lactobacillus kefir strains ATCC 35411 and ATCC 8007, but genotypic results failed to differentiate them (Mainville et al. 2006).
Genetic analysis of the embryo
Published in David K. Gardner, Ariel Weissman, Colin M. Howles, Zeev Shoham, Textbook of Assisted Reproductive Techniques, 2017
Yuval Yaron, Liran Hiersch, Veronica Gold, Sagit Peleg-Schalka, Mira Malcov
As an example, the ZFX and ZFY genes located on the X and Y chromosomes, respectively, can be distinguished according to differences in the size of the fragments pro- duced by the restriction enzyme HaeIII. This allows sex determination to be performed more accurately than that based on the presence or lack of amplification of the Y chromosome-specific SRY gene.
Association of NOD1, NOD2, PYDC1 and PYDC2 genes with Behcet’s disease susceptibility and clinical manifestations
Published in Ophthalmic Genetics, 2021
Ayca Kocaaga, Gunes Cakmak Genc, Sevim Karakas Celık, Rafet Koca, Ahmet Dursun
Genomic DNA was extracted from peripheral blood leucocytes by using E.Z.N.A® Blood DNA extraction kit according to the manufacturer’s instructions. rs2075820 (NOD1 G/A), rs2075818 (NOD1 G/C) gene polymorphisms and NOD2 R334Q/R334W gene mutations were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Amplifications were performed in 0.2 mL thinwall tubes of 50 μL aliquots containing 50 mM KCl, 10 mM Tris/HCI, 1.5 mM MgCl2, 0.5 μM of each of the four deoxynucleotides, 50 pmol of each primers, 1 U of Taq DNA polymerase (Bioron, cat. number: 101005) and 20 ng genomic DNA. The primer sequences, annealing temperatures, restriction enzymes and cleavage temperatures were used for SNPs analysis are given in Table 2. AvaI, HaeIII (BsuRI) and MspI (ThermoFisher S. Catalog numbers respectively: ER0381, ER0151 and ER0541) were used as restriction enzymes and reactions were performed in the “MWG primus thermal cycler-Primus 96 PCR system.”
Association of aldosterone synthase CYP11B2 (-344C/T) gene polymorphism with essential hypertension and left ventricular hypertrophy in the Egyptian population
Published in Clinical and Experimental Hypertension, 2019
The aldosterone synthase gene CYP11B2 −334C/T polymorphism was detected by PCR/RFLF as 1 µg of PCR product (15 µl) was added in a 25 µL reaction mixture containing 1 U (0.1 µl) of HAEIII restriction enzyme supplied by Roche Diagnostics GmbH, Mannheim, Germany, Catalog #: 10 693 944, 001, 2.5 µl of 10X SuRi/Cut buffer M and nuclease-free water and incubated at 37°C for 1 h then HAEIII restriction enzyme was thermally inactivated by incubation at 65°C for 15 min according to the manufacturer protocol. The digested PCR product was visualized after agarose gel electrophoresis by ultraviolet transillumination. The PCR product of 639 bp was subsequently cleaved by HaeIII, creating fragments for allele T 402, 138, 51 and 48 bp, and for allele C 334, 138, 68, 51 and 48 bp (Figure 2).
Sexually transmitted infections in oral cavity lesions: Human papillomavirus, Chlamydia trachomatis, and Herpes simplex virus
Published in Journal of Oral Microbiology, 2019
Jessica P. Mosmann, Angel D. Talavera, María I. Criscuolo, Raúl F. Venezuela, Ana X. Kiguen, Rene Panico, Ruth Ferreyra De Prato, Silvia A. López De Blanc, Viviana ré, Cecilia G. Cuffini
Viral and bacterial DNA was extracted using the commercial AccuPrepGenomic DNA Extraction Kit-Bioneer, following the manufacturer’s instructions. Then, HPV L1 genomic region (450 bp) was amplified with degenerate primers MY09 and MY11 following the protocol previously described by Manos et al. [11]. HPV-DNA positive samples were typed by restriction fragment length polymorphism (RFLP) method, using seven different restriction enzymes (BamHI, DdeI, HaeIII, HinfI, PstI, RsaI, and Sau3AIII) [12]. C.trachomatis was detected using CTP1 and CTP2 primers to cryptic plasmid obtained amplicons size of 201 bp [13]. We used a PCR with B3/B4 primer set designed to amplify a genomic fragment of highly conserved and divergent DNA sequences in the gene encoding the glycoprotein B (Gb) of primate α-herpesvirus. Then, RFLP with HaeIII restriction enzyme was used to differential identification of HSV-1 (555 bp) and HSV −2 (561 bp) [14].