scholarly journals HOT PHENOL EXTRACTION OF TOTAL RNA FROM Thermoascus aurantiacus AND CHARACTERIZATION OF ITS THERMOSTABLE XYLANASE GENE

2016 ◽  
Vol 1 (1) ◽  
pp. 55-58
Author(s):  
Ang Chung Huap ◽  
Awang Ahmad Sallehin Awang Husaini ◽  
Hairul Azman Roslan
Keyword(s):  
Reverse Transcriptase ◽  
28S Rrna ◽  
Thermoascus Aurantiacus ◽  
Xylanase Gene ◽  
Base Pairs ◽  
Total Rna ◽  
Phenol Extraction ◽  
Pcr Product ◽  
Polymerase Chain

Total RNA was successfully isolated using hot phenol extraction method. Three bands representing the 18S, 5.8S and 28S rRNA was observed. No heavy smearing was observed in the RNA band patterns, indicating low levels of polysaccharide contamination, when subjected to 1% agarose gel electrophoresis. Genomic DNA was eliminated using DNase I digestion and lithium chloride (LiCl) precipitation. Two-steps reverse transcriptase polymerase chain reaction (RT-PCR) using M-MuLV Reverse Transcriptase and sequence specific primers for xylanase gene, XynA(F) and XynA(R), successfully generated the target amplicon of 500 base pairs (bp). Sequence analysis of the PCR product indicated as partial sequence of Thermoascus aurantiacus xylanase gene (XynA) deposited in the NCBI GenBank with accession number: AF127529.1 and AJ132635.1. Hot phenol extraction is useful for extracting large quantities of total RNA sufficient for complementary DNA (cDNA) synthesis in shorter period of time.

scholarly journals Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran

2021 ◽  
Vol 9 (3) ◽  
pp. 257-265
Author(s):  
S. R. Naddaf ◽  
M. A. Oshaghi ◽  
H. Vatandoost ◽  
M. Assmar
Keyword(s):  
Sequence Data ◽  
Islamic Republic ◽  
Anopheles Fluviatilis ◽  
As Species ◽  
Pcr Product ◽  
Islamic Republic Of Iran ◽  
Polymerase Chain ◽  
Second Internal Transcribed Spacer ◽  
Species Specific

A species-specific polymerase chain reaction [PCR] assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran

Cultural and molecular characterization of photobionts of Peltigera Membranacea

1997 ◽  
Vol 29 (6) ◽  
pp. 571-586 ◽  
Author(s):  
V.P.W. Mioa ◽  
A. Rabenau ◽  
A. Lee
Keyword(s):  
16S Rdna ◽  
Ribosomal Rna ◽  
Molecular Study ◽  
16S Ribosomal Rna ◽  
Chain Reaction ◽  
Ribosomal Rna Gene ◽  
Pcr Product ◽  
16S Ribosomal Rna Gene ◽  
Polymerase Chain

AbstractA molecular study was undertaken to clarify the identity of the photobiont in colourmorphs of the lichen, Peltigera membranacea. Two strains of cyanobacteria, identified as Nosroc sp. by morphology, were cultivated from each of two lichen specimens. Prokaryotic (16S) ribosomal RNA gene fragments were amplified by the polymerase chain reaction (PCR) from DNA extracted from the isolated strains and the lichens, and sequenced directly. Sequences were 98 1% identical between lichen specimens, TDI#AR94 and TDI#AR95, and highly similar to sequences published, or generated in this study from a type culture, for Nostoc. The 16S ribosomal RNA gene sequences (‘ 16S rDNA’) of all four lichen-derived cyanobacteria appeared the same, even though the lichen specimens from which they originated had different sequences. The 16S rDXA from strains 9A and 9B were different from that of specimen TDI#AR94, the thallus from which they were isolated, and instead were the same as that of strains 10A and 10B, and their source, specimen TDI#AR95. When primers selective for the strain 9A sequence were used, however, a small amount of PCR product corresponding to the 16S rDNA of strain 9A was obtained from lichen TDI#AR94. The results confirm that the photobionts of P. membranacea belong to Nostoc, and suggest that genetic differences in the photobiont may be a factor in the occurrence of colourmorphs among cyanolichens.

Characterization of genetic polymorphism in the goat β-lactoglobulin gene

2000 ◽  
Vol 67 (2) ◽  
pp. 217-224 ◽  
Author(s):  
RAMONA N. PENA ◽  
ARMAND SÁNCHEZ ◽  
JOSEP M. FOLCH
Keyword(s):  
Fragment Length Polymorphism ◽  
Gene Frequencies ◽  
Single Nucleotide ◽  
Pcr Product ◽  
Polymorphic Position ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Β Lactoglobulin ◽  
New Alleles

Two new variants have been detected and characterized for the goat β-lactoglobulin gene at the cDNA level and confirmed at the genomic level. The two polymorphisms are located on exon 7 of the gene. One of the polymorphic sites is produced by a single nucleotide substitution in position +4601, allowing a polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) genotyping procedure to be developed using SacII restriction enzyme. The other polymorphic position contains a 10 bp long insertion at position +4641 that can be detected by capillary electrophoresis of the PCR product amplified with a fluorescent primer. The association of these two polymorphisms was also investigated, resulting in the description of two new alleles. Both of these contained the point mutation at the SacII site, with or without the 10 bp insertion at position +4641. The distribution of these new polymorphisms was studied in a population of males of four different goat breeds. The gene frequencies for these variants were similar in Spanish and French breeds.

Detection by reverse transcriptase-polymerase chain reaction and molecular characterization of subtype B avian metapneumovirus isolated in Brazil

2007 ◽  
Vol 36 (5) ◽  
pp. 383-387 ◽  
Author(s):  
Jorge Luis Chacón ◽  
Paulo E. Brandão ◽  
Marcos Buim ◽  
Laura Villarreal ◽  
Antonio J. Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Molecular Characterization ◽  
Chain Reaction ◽  
Avian Metapneumovirus ◽  
Subtype B ◽  
Polymerase Chain

scholarly journals Research Note. Trichinella spiralis parasitizing Puma concolor: first record in wildlife in Chile

2015 ◽  
Vol 52 (4) ◽  
pp. 360-363 ◽  
Author(s):  
C. Landaeta-Aqueveque ◽  
S. Krivokapich ◽  
G. M. Gatti ◽  
C. Gonzalez Prous ◽  
V. Rivera-Bückle ◽  
...  
Keyword(s):  
Puma Concolor ◽  
Trichinella Spiralis ◽  
First Record ◽  
Base Pairs ◽  
Pcr Product ◽  
Artificial Digestion ◽  
Polymerase Chain ◽  
Genotypic Identification

Summary The genus Trichinella is widespread in all continents but Antarctica. The only way to identify the species/genotypes is through molecular analyses. In Chile, only one study has reported Trichinella larvae in a cougar, but the species of Trichinella was not identified. In this work, the finding of Trichinella larvae in a cougar, together with their genotypic identification, is the first documentation of such in Chile. The cougar was found run over by a vehicle in the Biobío Region. Larvae were isolated following artificial digestion of the diaphragm and analyzed by means of multiplex polymerase chain reaction (PCR). A PCR product of 173 base pairs allowed for the classification of the larvae as T. spiralis. It is the first record of the species in Chilean wildlife. This finding in Chile is interesting in terms of human health, suggesting a possible role of the cougar as a reservoir for this parasite.

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