RNA isolation for transcriptomics of human and mouse small skin biopsies

<p>Abstract</p> <p>Background</p> <p>Isolation of RNA from skin biopsies presents a challenge, due to the tough nature of skin tissue and a high presence of RNases. As we lacked the dedicated equipment, i.e. homogenizer or bead-beater, needed for the available RNA from skin isolation methods, we adapted and tested our zebrafish single-embryo RNA-isolation protocol for RNA isolation from skin punch biopsies.</p> <p>Findings</p> <p>We tested our new RNA-isolation protocol in two experiments: a large-scale study with 97 human skin samples, and a small study with 16 mouse skin samples. Human skin was sampled with 4.0 mm biopsy punches and for the mouse skin different punch diameter sizes were tested; 1.0, 1.5, 2.0, and 2.5 mm. The average RNA yield in human samples was 1.5 μg with an average RNA quality RIN value of 8.1. For the mouse biopsies, the average RNA yield was 2.4 μg with an average RIN value of 7.5. For 96% of the human biopsies and 100% of the mouse biopsies we obtained enough high-quality RNA. The RNA samples were successfully tested in a transcriptomics analysis using the Affymetrix and Roche NimbleGen platforms.</p> <p>Conclusions</p> <p>Using our new RNA-isolation protocol, we were able to consistently isolate high-quality RNA, which is apt for further transcriptomics analysis. Furthermore, this method is already useable on biopsy material obtained with a punch diameter as small as 1.5 mm.</p

A comprehensive and synthetic dataset for global, regional, and national greenhouse gas emissions by sector

To track progress towards keeping global warming well below 2 ºC or even 1.5 °C, as agreed in the Paris Agreement, comprehensive up-to-date and reliable information on anthropogenic emissions and removals of greenhouse gas (GHG) emissions is required. Here we compile a new synthetic dataset on anthropogenic GHG emissions for 1970–2018 with a fast-track extension to 2019. Our dataset is global in coverage and includes CO2 emissions, CH4 emissions, N2O emissions, as well as those from fluorinated gases (F-gases: HFCs, PFCs, SF6, NF3) and provides country and sector details. We build this dataset from the version 6 release of the Emissions Database for Global Atmospheric Research (EDGAR v6) and three bookkeeping models for CO2 emissions from land use, land-use change, and forestry (LULUCF). We assess the uncertainties of global greenhouse gases at the 90 % confidence interval (5th–95th percentile range) by combining statistical analysis and comparisons of global emissions inventories and top-down atmospheric measurements with an expert judgement informed by the relevant scientific literature. We identify important data gaps for F-gas emissions. The agreement between our bottom-up inventory estimates and top-down atmospheric-based emissions estimates is relatively close for some F-gas species (∼ 10 % or less), but estimates can differ by an order of magnitude or more for others. Our aggregated F-gas estimate is about 10 % lower than top-down estimates in recent years. However, emissions from excluded F-gas species such as chlorofluorocarbons (CFCs) or hydrochlorofluorocarbons (HCFCs) are cumulatively larger than the sum of the reported species. Using global warming potential values with a 100-year time horizon from the Sixth Assessment Report by the Intergovernmental Panel on Climate Change (IPCC), global GHG emissions in 2018 amounted to 58 ± 6.1 GtCO2 eq. consisting of CO2 from fossil fuel combustion and industry (FFI) 38 ± 3.0 GtCO2, CO2-LULUCF 5.7 ± 4.0 GtCO2, CH4 10 ± 3.1 GtCO2 eq., N2O 2.6 ± 1.6 GtCO2 eq., and F-gases 1.3 ± 0.40 GtCO2 eq. Initial estimates suggest further growth of 1.3 GtCO2 eq. in GHG emissions to reach 59 ± 6.6 GtCO2 eq. by 2019. Our analysis of global trends in anthropogenic GHG emissions over the past 5 decades (1970–2018) highlights a pattern of varied but sustained emissions growth. There is high confidence that global anthropogenic GHG emissions have increased every decade, and emissions growth has been persistent across the different (groups of) gases. There is also high confidence that global anthropogenic GHG emissions levels were higher in 2009–2018 than in any previous decade and that GHG emissions levels grew throughout the most recent decade. While the average annual GHG emissions growth rate slowed between 2009 and 2018 (1.2 % yr−1) compared to 2000–2009 (2.4 % yr−1), the absolute increase in average annual GHG emissions by decade was never larger than between 2000–2009 and 2009–2018. Our analysis further reveals that there are no global sectors that show sustained reductions in GHG emissions. There are a number of countries that have reduced GHG emissions over the past decade, but these reductions are comparatively modest and outgrown by much larger emissions growth in some developing countries such as China, India, and Indonesia. There is a need to further develop independent, robust, and timely emissions estimates across all gases. As such, tracking progress in climate policy requires substantial investments in independent GHG emissions accounting and monitoring as well as in national and international statistical infrastructures. The data associated with this article (Minx et al., 2021) can be found at https://doi.org/10.5281/zenodo.5566761

A Re-Evaluation of the nuclear Structure Function Ratios for D, He, Li, C and Ca

We present a re-evaluation of the structure function ratios F2(He)/F2(D), F2(C)/F2(D) and F2(Ca)/F2(D) measured in deep inelastic muon-nucleus scattering at an incident muon momentum of 200 GeV. We also present the ratios F2(C)/F2(Li), F2(Ca)/F2(Li) and F2(Ca)/F2(C) measured at 90 GeV. The results are based on data already published by NMC; the main difference in the analysis is a correction for the masses of the deuterium targets and an improvement in the radiative corrections. The kinematic range covered is 0.0035 < x < 0.65, 0.5 < Q^2 <90 GeV^2 for the He/D, C/D and Ca/D data and 0.0085 < x < 0.6, 0.84 < Q^2 < 17 GeV^2 for the Li/C/Ca ones.Comment: 6 pages, Latex, 3 figures as uuencoded compressed tar file included at the end, in case of problems contact [email protected] (Antje Bruell