RAPD-PCR Analysis Research Articles

A preliminary Study on Genetic Diversity Among Different Genotypes of Manfalouty Pomegranate Cultivar B- Molecular Genetics Identification Using RAPD Markers

.), the most distributed commercial cultivar in Egypt, was investigated at a private farm located in El-Badari – Assuit governorate, Egypt. A preliminary surveys for 5 years in the field among Manfalouty cultivar was done and resulted in the selection of four promising pomegranate genotypes. Those genotypes proved to have desired characteristics and were given different names by local farmers i.e. (1) Abo- Shelh, (2) Almekhtat, (3) Abo-Shoka, and (4) Genah – Aldabor. The selected genotypes were characterized with molecular genetics identification using RAPD-PCR analysis. Polymorphism was detected among the selected genotypes by RAPD primers using five arbitrary primers for the four selected genotypes in comparison with Manfalouty cultivar. The percentage of polymorphism revealed by different primers ranged from 50% to 87.5% with an average of 64.44%. The UPGMA analysis of genetic distance of the four selected pomegranate genotypes and Manfalouty cultivar showed that, the highest value of similarity index was (0.86) occurred between Manfalouty cultivar and genotype (no.2) while the lowest value of similarity index was (0.67) occurred between the cultivars genotype (no.2) and genotype (no.4). From this study, we confirmed that diversity among the fruit characteristics by indication of genetic relatedness, while molecular tools are valuable to study such similarities.

Clonal diversity and antimicrobial resistance of Enterococcus faecalis isolated from endodontic infections

article i nfo Background: Enterococcus faecalis is considered to be one of most prevalent species in the oral cavity, particularly in endodontic infections. The aim of the present study was to investigate the prevalence of E. faecalis in dental root canals, clonal diversity by restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD-PCR) analysis, and the antibiotic susceptibility of E. faecalis isolates. Results: Among the bacterial strains isolated from dental root canal specimens (n = 82), E. faecalis was determined to have the highest prevalence followed by Streptococcus viridians, Leuconostoc mesenteroides, Staphylococcus aureus, Streptococcus mitis ,a ndPediococcus pentosaceus. Cluster analysis of RAPD-PCR and RFLP patterns of the E. faecalis isolates discriminated five and six different genotypes, respectively. Among the tested strains, 43%, 52% and 5% were susceptible, intermediate resistant, and resistant to erythromycin, respectively. In addition, one strain (E-12) was intermediate resistant to linezolid, and one isolate (E-16) was resistant to tetracycline. Interestingly, many of the intermediate resistant/resistant strains were grouped in clusters 5 and 6, according RAPD and to RFLP, respectively. Conclusions: E. faecalis demonstrated the highest prevalence in the tested dental root canal specimens collected from Saudi patients and were grouped into five to six different genotypes. Different levels of antimicrobial susceptibility were observed in the tested E. faecalis strains, which clearly indicated that although bacterial strains may be similar, point mutations can result in extreme susceptibility or resistance to various antibiotics. This phenomenon is a cause for concern for clinicians in the treatment of dental infections caused by E. faecalis.

English

Among the fungal diseases infecting tomato crops, early blight caused by Alternaria solani (Ellis and Martin) Jones and Grout is one of the most catastrophic disease causing accountable losses. Further, all of the tomato cultivars presently under cultivation have succumb more or less to early blight disease. Therefore, the present studies were undertaken for the pathogenic, cultural, morphological and molecular variability among the isolates of A. solani. The results reveal all of the eight isolates of A. solani as pathogenic to tomato (Cv. Pusa Ruby) and showed variability amongst them. The test isolates could grow better on the basic culture medium potato dextrose agar; however, highest mycelial growth was recorded on the isolate AsLt (88.50 mm), followed by AsBd (82.36 mm) and AsHl (78.40 mm), with excellent sporulation. All of the eight test isolates exhibited a wide range of variability in respect of their mycelial and conidial dimensions and septation. RAPD-PCR analysis of the four most virulent A. solani isolates, using 13 OPA primers revealed that the isolates AsBd (Beed) and AsLt (Latur) were closely related with 85% genetic similarity whereas, the isolates AsHl (Hingoli) and AsJl (Jalna) were closely related with 50% genetic similarity, but distinct from that of AsLt and AsBd isolates. Key words: Tomato, Alternaria solani, isolates, pathogenic, molecular variability, virulent, primers.

Functional diversity of phosphate solubilizing plant growth promoting rhizobacteria isolated from apple trees in the Trans Himalayan region of Himachal Pradesh, India

A total of 206 phosphate solubilizing bacteria (PSBs) were isolated from rhizosphere soil (RS) (116) and endorhizosphere (90) of apple trees of four locations: Chamba, Shimla, Kinnaur and Kullu of Himachal Pradesh, India. The population count of PSBs was invariably higher in rhizosphere than in root endosphere (ER) of apple. The greatest Shannon diversity indices for PSBs from RS (1.77) and ER (1.69) were obtained from Kinnaur. Polymorphism among 85 PSBs was detected using PCR-RAPD analysis. Sixteen representative PSBs from RAPD groups were identified further by biochemical tests and 16S rRNA gene analysis. Bacillus was the dominant genus represented by six different species, B. subtilis and B. licheniformis followed by B. pumilus, B. methylotrophicus, B. firmus and B.altitudinis suggesting diversity among species of the genus Bacillus. Variation exists in RS and ER PSBs suggesting the hypothesis that root exudates of apple plant exert selective pressure on the proliferation of specific kind of rhizobacteria on the surface of the root as well as within the root. B. methylotrophicus SMa5A, B. pumilus ECS10A and B. altitudinis ECH8A from ER and B. firmus KR4A from RS have been reported for the first time as plant growth promoting rhizobacteria from apple trees.

Spread of an Enterococcus faecalis sequence type 6 (CC2) clone in patients undergoing selective decontamination of the digestive tract.

Enterococcus faecalis (E. faecalis) is a common cause of nosocomial infection in immunocompromised patients. The presence and dissemination of high-risk clonal complexes, such as CC2, is an ongoing problem in hospitals. The aim of this work was to characterize 24 E. faecalis isolates from ICU patients undergoing selective decontamination of the digestive tract (SDD) by phenotypical (antimicrobial susceptibility) and genotypical (presence of virulence genes, RAPD-PCR and MLST) methods. Our results showed high prevalence of the ST6 E. faecalis clone (91.6%), especially adapted to the hospital environment, with a multidrug resistance pattern and a multitude of putative virulence genes. In addition, ST179 (4.2%) and ST191 (4.2%) were detected. By RAPD-PCR analysis, the 22 isolates identified as ST6 showed six different DNA patterns, while the two remaining isolates, ST179 and ST191, showed two additional profiles. CC2 is a known clonal complex with high adaptability to hospital environment and worldwide distribution. The high prevalence of the ST6 clone in the studied population could be related to the presence of gentamicin in the SDD mixture since most strains were gentamicin resistant. Consequently, strict surveillance should be applied for rapid detection and control of this clone to prevent future spread outside the ICU.

Morphological and Molecular Characterization ofFusariumspp. causing Post Flowering Stalk Rot of Maize

Ten isolates of Fusarium spp. (F. proliferatum (4 isolates), F. verticillioides (5 isolates), and F. pallidoroseum (1 isolates) were collected from different parts of Rajasthan from rotted maize stalks and identified by ITCC, IARI (New Delhi, India). The maximum colony diameter (90mm), sporulation (3.4x105 X 3.8x105), size of macroconidia (40.24 X 6.60) and microconidia (10.33 X 1.50) and septation (5–7) were observed in isolate Fv-01 (F. verticillioides) whereas minimum colony diameter (54.70mm), sporulation (0.8 x105 X 1.6 x105), size of macroconidia (17.57 X 1.80) and microconidia (9.09 X 1.48) was observed in isolate Fv-09 (F. proliferatum). On the basis of cultural and morphological characters of all the ten isolates showed variability, but based on RAPD-PCR analysis, these isolates can be categorized into two major clusters I and II at 0.44% similarity coefficient. RAPD-PCR analysis suggested that variability was observed between species of Fusarium recovered from maize and the technique could be use to complement morphological characterization and to determine genetic relationships between the species.

DNA fingerprinting and RAPD-PCR analysis ofS. aureusstrains isolated from bovine origin

DNA fingerprinting and RAPD-PCR analysis of<i>S. aureus</i>strains isolated from bovine origin

Naturalised populations of mesorhizobia in chickpea (Cicer arietinum L.) cropping soils: effects on nodule occupancy and productivity of commercial chickpea

Chickpea rhizobia did not occur naturally in Australian cropping soils, necessitating inoculation at sowing. Now, after more than 30 years of chickpea cultivation using a single inoculant strain, CC1192, it is likely that chickpea rhizobia are established in 1.0–1.5 Mha cropping land. The aims of this study were to examine effects of the naturalised chickpea rhizobia on nodulation and productivity (total crop N, crop N fixed and grain yield) of commercial chickpea. Soil was sampled from 26 fields to estimate chickpea rhizobial numbers, relate numbers to edaphic factors and years since previous chickpea crop, determine the proportions of CC1192 and novel strains using RAPD-PCR and subject a subset of novel strains from one site to 16S rRNA analysis. Nodules were harvested from 15 inoculated, commercial chickpea crops to determine occupancy by CC1192. The symbiotic effectiveness of a second subset of novel strains was assessed. The mean number of rhizobia in the soils varied from log 0.08 to log 5.16 rhizobia g soil−1 with population size positively correlated with soil moisture content and negatively correlated with salt concentration (ECe). RAPD-PCR analysis of 570 strains of chickpea rhizobia isolated from the soils indicated only 14 % with molecular fingerprints similar to CC1192. Occupancy by CC1192 of nodules harvested from commercial crops ranged 0–100 %, with an average of 53 %. Occupancy by CC1192 declined by an average 17 % with each log unit increase in numbers of novel chickpea rhizobia. We found no evidence that the novel mesorhizobia in the chickpea soils compromised N2 fixation or productivity of commercial chickpea crops, even though individual strains had generally reduced symbiotic effectiveness relative to CC1192.

Molecular characterization of Aspergillus infections in an Iranian educational hospital using RAPD-PCR method.

The nosocomial infections by Aspergillus species are associated with constructions and increased dust loads in hospital indoors. Our main object was to find the environmental sources of Aspergillus species causing hospital acquired infections. The clinical and environmental samplings were performed during 18 months from spring 2010 to summer 2011 in Imam educational hospital, Urmia, Iran. A morphological diagnosis was performed including microscopic characterization of isolated aspergillus from cultured specimens and polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) for the identification in the level of species. Random amplified polymorphic DNA - PCR RAPD-PCR using random primers for rDNA gene was performed to compare Aspergillus isolates of clinical cases with the relevant environmental sources. Use of RAPD method resulted various differential patterns, so that some Aspergillus isolates from the clinical and hospital indoor were completely matched (matched pairs) and some other Aspergillus isolates were not matched. In the case of matched pairs, Aspergillus niger and A. flavus isolated from broncoalveolar lavage and sinus discharge were relevant to those of air conditioner and walls surfaces, respectively. The hospital sources for the Aspergillus clinical isolates included air condition and walls. RAPD-PCR analysis can play a trivial role to find the hospital sources of Aspergillus clinical isolates.

Production of potential probiotic Spanish-style green table olives at pilot plant scale using multifunctional starters

This work evaluates the use of two multifunctional starters of Lactobacillus pentosus species (TOMC LAB2 and TOMC LAB4) during elaboration of Manzanilla olive fruits processed according to the Spanish-style. Data show that the use of inocula at the onset of fermentation led to a proper acidification and sugar consumption of brines compared to the spontaneous process, obtaining in a shorter period of time the maximum population for lactic acid bacteria. Both inoculated L. pentosus strains were recovered at high frequencies at the end of fermentation on the olive surface, which was corroborated by RAPD-PCR analysis. In situ observation of olive epidermis slices by scanning electron microscopy revealed a strong aggregation and adhesion between microorganisms, which reached population levels of approximately 6 and 7 log10 cfu/cm2 for yeasts and lactic acid bacteria, respectively. Enterobacteriaceae on the olive surface were also found at the onset of fermentation (∼9 log10 cfu/cm2), but they declined during the process and were below the detection limit at the end of fermentation. Results obtained in this study show the advantage of using multifunctional starters with the ability to adhere to the olive epidermis because, ultimately, the fruits are the food ingested by consumers.

Antifungal activity and identification of Lactobacilli, isolated from traditional dairy product “katak”

Filamentous moulds are the main spoilage microorganisms, responsible for significant economic losses and several healthy risks in human food chain. The lactic acid bacteria (LAB), especially lactobacilli could be a natural antagonist of these dangerous organisms. In Bulgaria, a very limited data exists on the antifungal activity of LAB microbiota of fermented dairy products. In the present study, four active strains were isolated from traditional fermented curd/yogurt-like product “katak”, produced in Bulgaria from centuries. The new isolates KR3, KR4, KR51 and KR53 were identified by API 50 CH biochemical test and different molecular methods (species-specific PCR, RAPD-PCR and 16S rDNA sequence analysis) as Lactobacillus brevis. According to our knowledge, this is the first data on the molecular characterization of the Lactobacillus microbiota of “katak”. A broad spectrum of antifungal activity of the four L. brevis KR strains against test-cultures representatives of carcinogenic, toxigenic, deteriorative and allergenic fungi from the genera Aspergillus, Fusarium, Penicillium and Trichoderma was estimated. Strains L. brevis KR3, KR4 and KR51 completely suppress the growth of Penicillium claviforme, Aspergillus awamori and Aspergillus niger. With regard to Aspergillus flavus and Trichoderma viride, a lower and strain-specific inhibitory activity was observed. The antifungal activity of our new L. brevis isolates seems to be a promising advantage of these four strains, suggesting their potential applications in different food technologies as bio-preservative agents against moulds.

Morphological-, chemical- and RAPD-PCR evaluation of eight different Ocimum basilicum L. gene bank accessions

In our study eight different Ocimum basilicum L. gene bank accessions were characterized by morphological evaluation, essential oil composition and RAPD molecular markers. Morphological traits were described using the standardized descriptor list of UPOV. The composition of the essential oil distilled from dried leaves was determined by gas chromatography/mass spectrometry (GC–MS). Fresh young leaves were used to extract genomic DNA for the RAPD-PCR evaluation. From the morphological aspects two groups could be distinguished with the method of cluster analysis. From the chemical aspect two different chemical types can be distinguished, according to the main components. The first one is a linalool type (‘Arvada’, ‘Dark Opal’, ‘Genovese’, ‘Lengyel’, ‘Rit-Sat’) and the second one is a linalool–methyl chavicol type (‘A-1’, Mittelgroβblättriger Grünes’, ‘Piros’). Based on RAPD-PCR analysis the accessions formed two clusters. The purple-leaved accessions (‘Piros’, ‘Dark Opal’) were located in group A, and the remaining ones (‘Genovese’, ‘Arvada’, ‘Lengyel’, ‘Rit-Sat’, ‘Mittelgroβblättriger Grünes’, ‘A-1′) were classified into group B. ‘Rit-Sat’ and ‘Genovese’ were not separated, based on their band pattern.It is shown by the result of the morphological, chemical and RAPD-PCR analysis that the data obtained by different methods not necessarily correlated with each other, except the accessions of ‘Genovese’ and ‘Dark Opal’, in which cases all characteristics show a good distinction. We observed that the cultivar ‘Genovese’ and the ‘Rit-Sat’ accession represent quite similar quality and the ‘Arvada’ accession of American origin is not really separated from the European ones by the complex evaluation.

RAPD-PCR and 16S rDNA phylogenetic analysis of alkaline protease producing bacteria isolated from soil of India: Identification and detection of genetic variability

bacterial strains were isolated from the soil samples collected from different regions of India out of which, 20 bacterial isolates were selected for alkaline protease production. The alka- line protease production efficiency of organisms was monitored at regular intervals (24 h) upto 7 days at 37 � C, pH 10. The 16S rDNA sequencing and RAPD-PCR based technique were used to identify the genetic variability among the 20 isolates of alkaline protease producing bacteria. The phylogenetic analysis indicated that the isolates can be separated into two clusters which could be further subdivided into five groups. Group 1 and 5 represented the family Bacillaceae, Groups 2 represented the Micrococcaceae family while Group 3 included the Arthrobacter bacterial group (family Micrococcaceae) from different geographical locations, respectively. Group 4 was identified as Pseudomonadaceae which was gram (� ) bacteria. 21 different oligonucleotide primers were used to amplify approximately 261 fragments from each DNA sample. The bands were scored on the basis of their presence and absence and similarity between DNA samples was checked using Jaccard's coefficient. Isolates were distinguished into distinct groups based on RAPD profiles from different geographical locations, morphological features and enzyme production efficiency. For cluster analysis the dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA). The results indicated that 16S rDNA and RAPD-PCR are suitable methods for rapid identification and differentiation of alkaline protease producing bacteria. a 2014 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology.

Molecular genetic structure of fruit fly (Bactrocera sp) population of South Andaman revealed through RAPD-PCR analysis

Fruit flies belonging to the family Dacinae, are biologically interesting and economically very important group of Diptera, as are known to infest a wide range of plant species especially fruits and vegetable crops. The present work was aimed for molecular profiling of fruit fly (Bactrocera sp) which is abundantly found in South Andaman. Though number of Bactrocera species has been reported to exist in these islands, Bactrocera cucurbitae is the most serious pest of cucurbit plants. Bactrocera sp is widely spread all over the world causing 85 -100 % damage to vegetable and fruits. Taxonomical diagnosis based on morphological characters could not discriminate them and all were found to be similar morphologically. During the course of this investigation, efforts were made to assess genetic similarities among these collections by using RAPD-PCR involving nine primers. The dendrogram of all primers depicted two major clusters which were having 60% similarity within as well as species. This shows a bright possibility of the use of RAPD-PCR in molecular profiling and identification of Bactrocera species of Andaman Islands.

Genetic variation of Chilo suppressalis Walker (Lepidoptera: Pyralidae) populations in Guilan and west of Mazandaran provinces analysed with RAPD markers

Genetic variation of striped stem borer&amp;nbsp;Chilo suppressalis&amp;nbsp;populations in Guilan and Mazandaran provinces in Iran was studied in 2010 by means of RAPD markers. Collected 45 samples from 19 locations (representing 17 counties in Guilan province and 2 counties in Mazandaran province) were classified into four groups of populations: three population groups corresponding to the west, centre, and east of Guilan and one population group from the west of Mazandaran. RAPD PCR analysis showed that&amp;nbsp;Chilo suppressalis&amp;nbsp;populations in Mazandaran are genetically different from the Guilan populations while the western populations of Guilan are genetically separated from the two other populations. The central and the eastern populations of Guilan exhibit some similarities with those of Mazandaran and the similarities are more obvious between the eastern populations of Guilan and the western population of Mazandaran. The population from the centre of Guilan showed the highest level of intra-population genetic variation possibly due to wider rice cultivated area and/or due to higher sample size. The results showed a high level of genetic variation in&amp;nbsp;Chilo suppressalis&amp;nbsp;populations in the two provinces in Iran. In spite of the model species is a widely distributed pest, the study indicated that the samples originated in populations with a different genetic make-up.

Prevalence and Molecular Diversity of Legionella pneumophila in Domestic Hot Water Systems of Private Apartments

Aims: To determine the prevalence of Legionella spp. in domestic hot water systems and evaluate the molecular diversity among these Legionella spp. Isolates. Place and Duration of Study: Sample collection area was the city of Aqaba, Jordan, between May and December 2012. Sample analysis was done in Ben-Hayyan international laboratories, Aqaba city, and the molecular microbiology laboratories, Taibah University, Saudi Arabia. Methodology: Two hundred (200) water samples were collected randomly from hot water tanks of private apartments, and were tested for the occurrence of Legionella spp. using direct membrane filtration method followed by species identification using Gram stain, the API 20NE biochemical system and the Legionella species latex agglutination test. Genotype characterizations of the Legionella isolates was carried out using DNA extraction followed by RAPD-PCR amplification with OP-A3 primer and analysis of the resulting patterns. Results: Of the 200 samples, 17 (8.5%) were positive for the presence of Legionella spp. A Original Research Article British Microbiology Research Journal, 4(3): 306-316, 2014 307 total of 15 (88.2%) out the 17 positive samples were confirmed as Legionella pneumophila, 10 of them were of serogroup 1 and 5 isolates were of serogroup 2-14, the remaining two isolate were Legionella species other than L. pneumophila. RAPD-PCR analysis classified all 17 Legionella isolates into three groups. Serogroup 1 isolates were classified into group A, serogroup 2-14 isolates in group B and Legionella spp. isolates in group C. Group A was further sub-clustered into two subgroups, genotype A1 containing isolates collected from hot water tanks of a temperature set at 25-30°C and A2 containing isolates collected from hot water tanks of a temperature set at 55-80°C. Conclusion: This study showed the colonization of the plumbing systems of private houses by Legionella spp. and demonstrated that the temperature of the water tanks maybe one of the most important factors that affect the genotypic behavior of Legionella pneumophila.

Molecular Studies on Salmonella Species Isolated From Chicken

Although many pathogens have recently received considerable attention, Salmonella remains among the leading sources of food-borne illness throughout much of the world. A poultry producer suffers losses due to Salmonella infection of the flock including loss of birds and production time. The present study was carried out to compare between the conventional bacteriological method and pre-enriched PCR assay for detection of Salmonella in chicken swabs samples The culture method can detect Salmonella in 6 samples out of 105 of examined samples under investigation with an incidence of 5.7 %, where by using PCR can detect 7 Salmonella positive samples out of 105 examined samples with an incidence of 6.7 %. Dendrogram or phylogentic tree of 5 isolated salmonella serotypes on RAPD-PCR analysis – PCR pattern similarity. The phylogentic tree was constructed applying UPGMA to matrix of similarity established from the combined data obtained by using three RAPD-PCR primers (1, 2, and primer 5) which give the best discrimination out of the six Primers used in the study. The numbers indicate the percentage of similarity among RAPD-PCR profiles based on pair wise band sharing as compared with the molecular weight of the marker (DNA ladder).. it was found that Salmonella serotypes isolated chickens and environmental samples have different degrees of genetic similarity as follow; S. anatum with S. typhimurium (52 %), S. anatum with S. derby (57 %), S. anatum with S. kentucky ( 62 %), S. anatum with S. enteritidis ( 48 %). S. typhimurium with S. derby ( 67 %), S. typhimurium with S. kentucky ( 43 %), S. typhimurium with S. enteritidis ( 38 %). S. derby with S. kentucky ( 37 %) S. derby with S. enteritidis (33 %). S. kentucky with S. enteritidis (29%). These results indicate that there are a different degrees of polymorphism between the 5 serotypes under investigation from lower degree of genetic similarity between (29%) between S. kentucky and S. enteritidis to higher degree (67 %) between S. typhimurium with S. derby.

Characterization of Fasciola gigantica isolates from cattle from South-western Zimbabwe using RAPD-PCR

This ia n journal article in the IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS), published by International Organization Of Scientific Research (IOSR)in 2014. It is focusing on the characterisation of fasciola gigantica isolates from cattle using RAPD-RCR. The study is based in Zimbabwe's Western region.,The study sought to characterize Fasciola gigantica isolates from cattle in different localities using RAPD-PCR. Adult flukes morphologically identified as F. gigantica were collected from slaughtered infected animals during meat hygiene inspections. DNA was extracted from single flukes and subjected to RAPD-PCR analysis. In the RAPD-PCR analysis, genomic DNA isolated from the conical anterior end of the worms was amplified by the polymerase chain reaction using 10 random oligonucleotide primers. Depending upon the Fasciola gigantica isolate-primer combination, 1-13 DNA fragments in the range of 75-2000bp were amplified. It was observed that all the 10 primers directing amplification of DNA were of potential interest in the generation of polymorphic DNA. The percentage polymorphic loci ranged from 33.33-100%. Polymorphic bands were scored and used to calculate Nei?s 1978 genetic distance. The genetic distance values ranged between 0.0690 (isolate 5 and 6 from Gwanda and 0.6109 (isolate 6 from Gwanda and isolate 14 from Matopo). The mean Nei?s gene diversity was 0.2839. The study showed the variability of Fasciola gigantica isolates from the same host, using RAPD markers could be applied as a low cost way of identification

Detection of Genomic Instability in Renal Cancer by Random Amplified Polymorphic DNA Analysis from Urine Sample as a Non-invasive Method: Potential Use in Diagnosis

The study applied random amplified polymorphic DNA (RAPD) analysis including PCR technique in detection of genomic instability of renal cancer from urine samples as an early detective and a noninvasive method. DNAs extracted, using manual method, from exfoliated cells in urine of 52 renal cancer patients. All yielded DNAs were amplified with the same 10-base pair random primer (BioA-09) with GC rich sequence (5’-GGGTAACGCC-3’). PCR products from RAPD analysis were electrophoretically separated in agarose gels and banding profiles were visualized by ethidium bromide staining and viewed under ultraviolet light. Genomic alteration has been shown and included the disappearing of normal bands and appearing of new bands related to the tumor. The correlation between the genomic alteration and the histopathological types was assessed by using the X 2 test. Bands with molecular sizes 180 (X 2 = 13.15, p <0.001), 250 (X 2 = 9.524, p = 0.002), and 1250(X 2 = 5.357, p = 0.021) bp can be utilized as molecular markers for conventional renal cell carcinoma in addition to 1700 bp utilized as molecular marker for transitional cell carcinoma of renal pelvis (P = 0.005). Also there is an association between the presence of 500 bp and renal tumor as general (90.4%). The results of current investigation support the availability of using of RAPD-PCR analysis for screening of specific molecular markers by using Bio-A09 primer. These markers might be useful in diagnosis of renal cancer types and inspecting the tumor development.

Composition of raw cow milk and artisanal yoghurt collected in Maroua (Cameroon)

This study attempts to identify species of Euphorbia ( Euphorbia peplus, Euphorbia helioscopia, Euphorbia granulata and Euphorbia hirta ) grown in University of Baghdad Campus in Jadiriyah and determine the genetic polymorphism among them by using DNA markers generated by polymerase chain reaction (PCR). Total genomic DNA of species studied was extracted from dry seeds by using commercial kit. Molecular analysis was performed by using nine random markers in random amplified polymorphic DNA (RAPD-PCR) technique. RAPD-PCR analyses based on three primers A13, C05 and D20 gave results in term of amplification and polymorphisim for the four species studied. The genetic polymorphisms value of each primer was determined and ranged between 47 to 84%; primer A13 produced the highest percent of genetic polymorphism compared with primer C05. RAPD-PCR technique confirmed the isolation of the four species of Euphorbia obviously. Key words: Euphorbia spp ., random markers in random amplified polymorphic DNA (RAPD-PCR), monomorphic, polymorphic, random primers.