Showing papers on "Restriction map published in 2019"
TL;DR: A proof of principle system was developed to elute and concentrate lambda DNA molecules at the interface between a solution and a poly‐acrylamide roadblock, slowing down the progression of DNA in the matrix, since the molecules were so large and the pore size was small.
Abstract: To understand structural variation for personal genomics, an extensive ensemble of large DNA molecules will be required to span large structural variations. Nanocoding, a whole-genome analysis platform, can analyze large DNA molecules for the construction of physical restriction maps of entire genomes. However, handling of large DNA is difficult and a system is needed to concentrate large DNA molecules, while keeping the molecules intact. Insert technology was developed to protect large DNA molecules during routine cell lysis and molecular biology techniques. However, eluting and concentrating DNA molecules has been difficult in the past. Utilizing 3D printed mesofluidic device, a proof of principle system was developed to elute and concentrate lambda DNA molecules at the interface between a solution and a poly-acrylamide roadblock. The matrix allowed buffer solution to move through the pores in the matrix; however, it slowed down the progression of DNA in the matrix, since the molecules were so large and the pore size was small. Using fluorescence intensity of the insert, 84% of DNA was eluted from the insert and 45% of DNA was recovered in solution from the eluted DNA. DNA recovered was digested with a restriction enzyme to determine that the DNA molecules remained full length during the elution and concentration of DNA.
2 citations
TL;DR: This study compared restriction map of hemagglutinin-neuraminidase gene encoding sialidase protein among Lentogenic strains and Velogenic isolates obtained in Iran, and a variety of virus genotypes retrieved from GenBank to suggest this method might be useful for pathotyping and differentiation of class I and II Newcastle disease virus.
Abstract: Newcastle disease virus (NDV) is considered as a deadly infection to majority of the avian species One of the key factors responsible for virus pathogenesis is sialidase protein expressed by hemagglutinin-neuraminidase gene on the envelope of virus particles, which involves in virus attachment to cells In this study we compared restriction map of hemagglutinin-neuraminidase gene encoding sialidase protein among Lentogenic strains and Velogenic isolates obtained in Iran, and a variety of virus genotypes (I–VII) retrieved from GenBank A polymorphic region with 527 bp length was found in the mentioned gene Reverse transcription-Polymerase chain reaction was done for Iranian Velogenic isolates and vaccine strains (B1, La Sota and V4) The Pattern of four restriction endonuclease enzymes (AluI, AvaIl, HaeIII and TaqI) was analyzed by Polymerase chain reaction-Restriction fragment length polymorphism technique There were significant differences among Iranian Velogenic isolates and vaccine strains The experimental results of the mentioned method were in concordance with in silico evaluations of restriction map of 146 strains including: class I and class II genotypes obtained from the National Center for Biotechnology Information (NCBI) Therefore, we suggest that this method might be useful for pathotyping and differentiation of class I and II Newcastle disease virus
2 citations
TL;DR: Measuring plasmid DNA diversity together with the antibiotic resistance indicates an epidemic transfer and acquisition of different plasmids in pathogenic E. coli as a result of antibiotic random treatment.
Abstract: Pathogenic bacteria are constantly adapted against antimicrobial drugs by arising new traits of drug resistance. The genes of such resistance are mostly possessed by pathogens due to the random use of antibiotics and commonly held on plasmid DNA. Plasmid-mediated antimicrobial resistance genes are easily transferred horizontally from one bacterial cell to another in an epidemic manner causing an increasing and serious challenge to clinicians to overcome the infectious pathogens. UTI is the most common infection caused by bacteria which has become hard to be treated due to the emerging problem of antibiotic resistance. The current study correlates plasmid DNA diversity and antibiotic resistance in E. coli; the common causative agent of UTI. Obtaining plasmid restriction maps provides a clear view about how DNA could be diversified in one bacteria. The result showed a clear DNA polymorphism in plasmids purified from E. coli. There has been 10 forms of different restriction plasmid profiles among 63 sample of E. coli. The 10 forms of plasmid profiles have been classified into two groups: highly diversified and lowly diversified profiles. Strains of highly diversified plasmid profiles showed significantly more resistance toward antibiotics than strains of less diversified plasmid profiles. Measuring plasmid DNA diversity together with the antibiotic resistance indicates an epidemic transfer and acquisition of different plasmids in pathogenic E. coli as a result of antibiotic random treatment.