Genetic variations among prawns act as an important tool to characterize and differentiate between the species. Molecular and phylogenetic analysis of shrimps and prawns like any other organism rely on high yields of pure and better quality genomic DNA. In this regard isolation of DNA is the first and basic step. In spite of the availability of many protocols of DNA extraction from animal tissues, it is difficult to ascertain that which one would provide desired results for prawn tissue. In the present study, three different techniques of DNA isolation i.e., salting out, phenol-chloroform and Qiagen DNA extraction kit were performed and compared for their yield. Cephalothoracic tissue and muscle tissue of pleopods were used for isolation. Tissue samples from fresh specimens as well as from alcohol preserved specimens were employed for extraction. The quantity (µg/ml) and quality of isolated DNA were determined by UV spectrophotometry and agarose gel electrophoresis. Results showed that Phenol-chloroform method with slight modifications obtained higher yield of genomic DNA as compared to other methods. The present work also revealed that among fresh specimens cephalotoracic tissue yielded high concentration DNA than muscle tissue. However, among alcohol preserved specimens, the concentration of DNA was higher in muscle tissue of pleopods. The high quality DNA was then subjected to randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) analysis. The DNAs produced clear, sharp and reproducible PCR (Polymerse chain reaction) product pattern.
Disturbance in vascular functioning pathways has been related to pathophysiology of migraine. The present study investigated the role of MTHFR C677T and ACE I/D gene polymorphisms in migraine susceptibility within the population of Jammu province of J&K state. A sum of 252 subjects including 102 migraine patients and 150 non-migrainous unrelated healthy controls were enrolled for the present study. PCR-RFLP was performed for determining MTHFR gene variations. For detecting insertion/deletion in ACE gene PCR was performed. In case of MTHFR, ‘T’ allele (variant allele) and TT genotype (variant) was found to be present only in migraine patients but not in controls thereby suggesting its positive role in migraine pathophysiology. For ACE I/D polymorphism, higher frequency of DD genotype (32.35 % vs 15.3 %) and D allele (0.51 vs 0.4) were observed in patients than in controls. Logistic regression analysis revealed a significant association of ACE I/D polymorphism with risk of migraine. However, a direct link of MTHFR C677T polymorphism with migraine risk was not found.
<b><i>Introduction:</i></b> Migraine is a neurovascular disorder and is clinically characterized by episodic attacks of mild to severe headaches. Due to the involvement of multiple environmental and genetic factors, it has become a much more complex neurological condition to understand. Apart from the environmental variables, a plethora of genes have been implicated, and one such example is <i>ESR</i>1. The present study was focused to find out the association of two important polymorphisms, namely, <i>Pvu</i>II and <i>Xba</i>I of the <i>ESR</i>1 with migraine in the population of Jammu and Kashmir (UT). <b><i>Methods:</i></b> The PCR-RFLP genotyping method was utilized to detect <i>Pvu</i>II and <i>Xba</i>I polymorphism, and the result was confirmed by statistical analysis. <b><i>Results:</i></b> Although we did not find a signification association of <i>ESR</i>-<i>Pvu</i>II polymorphism with migraine susceptibility {OR: 1.14 at 95% CI [0.76–1.71] (<i>p</i> value 0.5)}, a strong association was found with the clinical subtype of migraine; migraine with aura (MA) {OR: 2.014 at 95% CI [1.069–3.792] (<i>p</i> value 0.028)}. Furthermore, a significant association of <i>ESR</i>-<i>Xba</i>I polymorphism was observed with migraine {OR: 1.908 at 95% CI [1.252–2.907] (<i>p</i> value 0.002) and its both clinical subtypes; migraine without aura (MO) {OR: 1.870 at 95% CI [1.186–2.950] (<i>p</i> value 0.006)} and MA {OR: 2.014 at 95% CI [1.069–3.792] (<i>p</i> value 0.028)}. <b><i>Conclusion:</i></b> In conclusion, <i>ESR</i>1-<i>Xba</i>I polymorphism is significantly associated with migraine risk including both subtypes (MA and MO) in the North Indian population of Jammu.
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