Proper interpretation of RNA sequencing data requires an understanding of assay sensitivity and sources of variability. To this end, the External RNA Control Consortium (ERCC) developed a standard set of 92 poly-adenylated RNA transcripts that are orthogonal to mammalian RNA that can be added to RNA extracts before library generation and sequencing. The presence of these RNA standards at known ratios improves interpretation of RNA sequencing datasets. To test the utility of the ERCC RNA controls, total RNA extracted from mouse livers from the Rodent Research 1 (flight, ground control, basal control and vivarium control groups) and Rodent Research 3 (flight, ground control and basal control groups) missions were sequenced with and without the ERCC control RNA (RR-3 liver samples without ERCC control have been sequenced previously and the dataset is released in the GLDS-137). To allow comparison within and between groups, ERCC Mix 1 or Mix 2 were added to half of the samples from each group, respectively.

Supplementary data for "Comparison of whole transcriptome and targeted RNA sequencing for ecological high-throughput transcriptomics". This dataset is associated with the following publication: Villeneuve, D., M. Nash, A. Biales, K. Bush, G. Evensen, L. Everett, J. Haselman, M. Hazemi, M. Le, H. Poynton, B. Seligmann , L. Wehmas, J. Yeakley, and K. Flynn. Comparison of whole transcriptome and targeted RNA sequencing for ecological high-throughput transcriptomics. REGULATORY TOXICOLOGY AND PHARMACOLOGY. Elsevier Science Ltd, New York, NY, USA, 162: 105898, (2025).

,This dataset presents the Neodiprion Official Gene Set (OGS) v1.1. It was generated using Maker v2.31.8, followed by CrossMap re-mapping of coordinates to genome assembly Nlec1.1 (https://www.ncbi.nlm.nih.gov/assembly/GCA_001263575.2/).,This dataset is now obsolete - a new genome assembly, iyNeoleco1.1 (https://www.ncbi.nlm.nih.gov/assembly/GCF_021901455.1) has been produced by the Ag100Pest project, with annotations from NCBI's RefSeq resource.,,

Proper interpretation of RNA sequencing data requires an understanding of assay sensitivity and sources of variability. To this end the External RNA Control Consortium (ERCC) developed a standard set of 92 poly-adenylated RNA transcripts that are orthogonal to mammalian RNA that can be added to RNA extracts before library generation and sequencing. The presence of these RNA standards at known ratios improves interpretation of RNA sequencing datasets. To test the utility of the ERCC RNA controls total RNA extracted from mouse livers from the Rodent Research 1 (flight ground control basal control and vivarium control groups) and Rodent Research 3 (flight ground control and basal control groups) missions were sequenced with and without the ERCC control RNA (RR-3 liver samples without ERCC control have been sequenced previously and the dataset is released in the GLDS-137). To allow comparison within and between groups ERCC Mix 1 or Mix 2 were added to half of the samples from each group respectively.

The data contained in this worksheets provide sequences submitted for public access, analysis for RNA sequences generated in this study. The data and analysis are for a manuscript "The trait repertoire enabling cyanobacterial blooms assessed through comparative genomic complexity ". This dataset is associated with the following publication: Cao, H., Y. Shimura, M.M. Steffen, Z. Yang, J. Lu, A. Joel, L. Jenkins, M. Kawachi, Y. Yin, and F. Garcia-Pichel. The Trait Repertoire Enabling Cyanobacteria to Bloom Assessed through Comparative Genomic Complexity and Metatranscriptomics. mBio. American Society for Microbiology, Washington, DC, USA, 11(3): e01155-20, (2020).

NIST developed a reference material consisting of synthetic fragments of the SARS-CoV-2 virus RNA, which is the target of molecular diagnostic tests. These RNA fragments can assist in the development and validation of RT-qPCR assays for the detection SARS-CoV-2. The RNA fragments are characterized for concentration using digital PCR methods, may be used to assess limits of detection for SARS-CoV-2 assays, and may calibrate other in-house or commercial SARS-CoV-2 controls. This dataset includes data used for RTGM 10169 SARs-Cov-2 Research Grade Test Material validation. The Illumina and ONT sequence data were used to verify the construct sequence.

Background PCR amplification of target molecules involves sequence specific primers that flank the region to be amplified. While this technique is generally routine, its applicability may not be sufficient to generate a desired target molecule from two separate regions involving intron /exon boundaries. For these situations, the generation of full-length complementary DNAs from two partial genomic clones becomes necessary for the family of low abundance genes. Results The first approach we used for the isolation of full-length cDNA from two known genomic clones of Hox genes was based on fusion PCR. Here we describe a simple and efficient method of amplification for homeobox D13 (HOXD13) full length cDNA from two partial genomic clones. Specific 5' and 3' untranslated region (UTR) primer pairs and website program (primer3_www.cgv0.2) were key steps involved in this process. Conclusions We have devised a simple, rapid and easy method for generating cDNA clone from genomic sequences. The full length HOXD13 clone (1.1 kb) generated with this technique was confirmed by sequence analysis. This simple approach can be utilized to generate full-length cDNA clones from available partial genomic sequences.

Low RNA yield and quality limit use of formalin-fixed paraffin-embedded (FFPE) tissue samples for genomic analyses. In this study, we evaluated methods to demodify RNA highly fragmented and crosslinked by formalin fixation. Primary endpoints were RNA recovery, RNA-sequencing quality metrics, and target gene responses to a reference chemical (phenobarbital, PB). Frozen mouse liver samples from control and PB groups (n=6/group) were divided and preserved for 3 months as follows: frozen (FR); 70% ethanol (OH); 10% buffered formalin for 18 hours followed by ethanol (18F); and 10% buffered formalin (3F). Samples from OH, 18F, and 3F groups were processed to FFPE blocks and sectioned for RNA isolation. The latter group received no additional treatment (3F) or the following demodification protocols: short heated incubation with TAE buffer; overnight heated incubation with an organocatalyst using two different isolation kits; or overnight heated incubation without organocatalyst. TruSeq Stranded Total RNA libraries with Ribo-Zero were built and sequenced using the Illumina HiSeq platform. Extended incubation with or without organocatalyst increased RNA yield >3-fold and enhanced quality compared to 3F, as indicated by higher RNA integrity number (>1.5-fold) and fragment analysis values (>3.0-fold). Post-sequencing metrics showed reduced bias in gene coverage and deletion rates for all extended incubation groups. Following PB-induced differential gene expression analysis, all demodification groups showed increased overlap with FR in genes (73-83%) and pathways (91-94%) compared to 3F overlap with FR (60% and 63%, respectively). These results demonstrate simple changes in RNA isolation methods that can enhance genomic analyses of FFPE samples. This dataset is associated with the following publication: Wehmas, L., C. Wood, R. Gagne, A. Williams, C. Yauk, M. Gosink, D. Dalmas, R. Hao, R. O'Lone, and S. Hester. Demodifying RNA for Transcriptomic Analyses of Archival Formalin-Fixed Paraffin-Embedded Samples. TOXICOLOGICAL SCIENCES. Society of Toxicology, RESTON, VA, 162(2): 535-547, (2018).