Random non-coding fragments of lizard DNA: anonymous nuclear loci for the Australian skink, Tiliqua rugosa, and their utility in other Egernia-group species

We report the development of 48 anonymous nuclear loci from the Australian skink Tiliqua rugosa using 454 sequencing. These loci amplified across a Western Australian lineage (47 loci), a ‘northern’ lineage (48 loci) and a ‘southern’ lineage (46 loci). We further tested amplification for the related T. adelaidensis and Egernia stokesii where 37 and 34 loci amplified respectively. The loci showed variability within T. rugosa (22 polymorphic loci) and at least 27 loci also exhibited variation among the three species, highlighting the usefulness of these markers for phylogenetic, phylogeographic and population genetic analyses in T. rugosa and related species.

EARLY DIAGNOSIS OF HYPERTENSION WITH MOLECULAR GENETIC TECHNIQUES

Objective: To standardize the molecular techniques for early diagnosisof genes of hypertension. Methodology and Results: These techniques include extraction of DNA in which weextracted DNA by manual method with results of high yield, less purity and it was more time consuming. On theother hand with kit (Fermentas) method yield was low with high purity and less time consuming. The purity of DNAwas checked by spectrophotometer by using DNA/RNA ratio. Conditions for diagnosis was optimized for specificDNA sequence by using primers for genes Agt and Ace. Restriction digestion was done with the amplified productwith restriction enzymes Lwel and Nco1 the result was found negative with no polymorphism. Single strandedconformational polymorphism (SSCP) was performed which is more efficient method of obtaining information aboutlevel of polymorphism with in anonymous nuclear loci than the restriction enzyme protocol. SSCP is also morespecific because it gives idea which specific portion of gene is highly polymorphic. The result was again negative.Conclusion: This preliminary study of molecular analysis optimized the conditions for detection of polymorphism ofcandidate genes associated with hypertension. Following the same standardized conditions this is possible that wecan study and diagnose thousands of hypertensive patients. This is very much helpful for future planning of personswho are prone to hypertension due to family history.