Gene expression studies require high-quality intact total RNA for real-time PCR analysis. Degraded or impure sample can provide disastrous results in the subsequence process. RNA is very unstable and can be contaminated easily by ribonuclease (RNase), therefore utmost quality control checks are required to ensure best quality RNA with high yield. Herein, we have come out with a protocol to isolate total RNA from liver, kidney and brain of adult zebrafish using a commercially available chemical denaturant and subsequent clean-up to remove traces of DNA and impurities. Kidney tissue gives maximum yield of total RNA concentration i.e. 135.8786 ng/µL. Purity of isolated RNA was observed with an absorbance ranging from 1.9 to 2.0 observed in all samples. The whole procedure was repeatedly performed to get satisfactory results. Throughout these procedures there are numerous quality control checks to ensure that the sample is neither degraded nor contaminated.
Review Article IJOAR (2020) 3:68 Smokeless Tobacco dependence and cessation measures in India India being the largest smokeless tobacco (SLT) producer and consumer in the South East Asia Region (SEAR) entitled with a range of products. SLT consumption is very common because it is associated with socioeconomic and cultural perspectives in the India. After the implementation of the first treaty of the World Health Organization Framework Convention on Tobacco Control (WHO-FCTC), public health standards were reaffirmed and improvised through successful implementation of articles since 2007 as reported in the global progress factsheet. Many measures have been implemented for achieving the success of WHO-FCTC in India and to control the huge disease burden induced by tobacco use. The objective of the present study was to understand and explore the factors involved in the SLT demand reduction by revealing the dependence and cessation implementation in India in the context of strategies and policies. For this study, we searched for SLT dependence and SLT cessation literature survey in PubMed and obtained 102 literatures consisting of studies and reviews, which were further scrutinized by excluding the reviews, studies conducted abroad and studies conducted within the last 5 years (after 2015). No statistical significance was observed in comparison of National to International SLT dependence and an attempt to quit SLT following the tenure of a year. This is in-line with GATS-2 and international data. We have also discussed a diverse approach in the control of SLT at different levels-such as SLT users, healthcare professional, policy advocates supplier, labeling, legal policies, and educating at various platforms. Effective strategies for the cessation of SLT is mandatory with proper guidance for the manufacturer, users and sellers in controlling SLT products.
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