Notes: Researchers used the SV Total RNA Isolation System to isolate total RNA from various tissues from Drosophila larvae. The isolated RNA from various tissues was used in reverse transcription reactions (using M-MLV Reverse Transcriptase) to make cDNAs of the shade (Shd) gene message. PCR amplification of Shd cDNA demonstrated that Shd was specifically expressed in certain Drosophila tissues. (2784)

Notes: The species-specific response of mouse hepatoma (Hepa.2D) and guinea pig adenocarcinoma (GPC.16) cells to flavonoid treatment was explored in this study. Cell lines stably transfected with dioxin responsive element-driven luciferase constructs were treated with a series of flavanoids. Luciferase expression was measured using the Steady-Glo®> Luciferase Assay system. To increase the number of cells available, cells were grown on Cytodex-1 beads in a larger volume, then split into an opaque 96-well plate prior to application of the Steady-Glo® Reagent. Total RNA was isolated using the SV Total RNA Isolation System, then used in real-time PCR to quantitate luciferase and GAPDH RNA levels after treatment. (3091)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from Tobacco Hornworn (M. Sexta) larvae. The RNA was reverse transcribed with primers specific to the δ-endotoxin-binding region from the cadherin ectodomain of BT-R1. (2785)

Notes: To analyze whether the presence or absence of calcium in Yersinia enterocolitica growth media affected secretion of Yersinia outer proteins (Yops), these researchers used mutational analysis on four gene products that work together to regulate yopQ expression. The various Y. enterocolitica strains were grown in TSB medium supplemented with 5mM CaCl₂ or EGTA (+/-Ca²⁺). After a 2-hour incubation followed by a 2-hour induction, the cultures were centrifuged and the supernatant was separated from the cell pellet. After resuspension in cell lysis buffer, the pellet was subjected to French press and 100µl aliquots were used with the SV Total RNA Isolation System. The eluted total RNA was digested with DNase, phenol/chloroform extracted and ethanol precipitated. Once resuspended, 0.6µg of Yersinia RNA was used in two-step RT-PCR to amplify the yopQ gene and the amplimer was analyzed on an ethidium bromide-stained 2% agarose gel. (3071)

Notes: The authors describe using the Wizard^® Genomic DNA Purification kit and the SV Total RNA Isolation System to isolate genomic DNA and total RNA, respectively, from Streptococcus pneumoniae. The researchers also added RNasin^® Ribonuclease Inhibitor to purified total RNA before storing it and using it for later RT-PCR reactions performed with the Access RT-PCR System. (2835)

Notes: In this paper, the SV Total RNA Isolation System was used to isolate total RNA from Gigaspora margarita. The authors reported isolating total RNA from 100 spores or 100mg of mycorrhizal roots. One microliter of the isolated RNA was used in RT-PCR to amplify Gigaspora margarita metallothionein (MT)-like polypeptide (GmarMT1) RNA. The researchers also cloned the GmarMT1 coding region using the pGEM^®-T Vector. (3077)

Notes: Total RNA was isolated from 100mg of ferret aorta tissue using the SV Total RNA Isolation System. One microgram of the purified total RNA was used in RT-PCR to amplify cDNA of CaMKII mRNA. The PCR products were cloned and sequenced. (2851)

Notes: Total RNA was isolated from TOP10F’ or INVaF' E. coli (Invitrogen) using the SV Total RNA Isolation System. The isolated RNA was used in RT-PCR to detect cleavage of marA mRNA. A marA drug resistance gene sequence was also ligated into a chloramphenicol acetyltransferase (CAT) reporter vector. This construct was used to measure single-stranded nanovector-transcribed ribozyme activity on the marA sequence. CAT activity was measured using the CAT Enzyme Assay System. CAT activity was normalized by cotransfecting cells with the pSV-β-Galactosidase Vector and assaying lysates with the β-Galactosidase Enzyme Assay System. (2794)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from vastus lateralis muscle biopsies from 20 male and female subjects before and after a nine-week strength-training regime. One microgram of pooled total RNA from each age/sex group was then reverse transcribed to make ³³P-labeled cDNAs. Labeled cDNAs from each group were hybridized to GF211 Microarrays (Invitrogen) and analyzed for differential expression patterns. (2759)

Notes: Total RNA was isolated from taproots, etiolated heads, green leaves, and green and etiolated seedlings of chicory using the SV Total RNA Isolation System. Two micrograms of RNA from each sample were used in a Northern blot to analyze gene expression. (2854)

Notes: Total RNA was isolated from Microcystis aeruginosa for use in RT-PCR and RACE (Rapid Amplification of cDNA Ends) to characterize transcripts of a gene cluster encoding microcystin synthetase. (2311)

Notes: The authors used the SV Total RNA Isolation System to purify total RNA from 1 ml of whole blood anticoagulated with EDTA for RT-PCR studies of prostate-specific antigens. (2470)

Notes: The SV Total RNA Isolation System was used to isolated RNA from various dissected mouse brain areas. Ten microliters of the isolated RNA were used for RT-PCR analysis of gene expression patterns. (2186)

Notes: The SV Total RNA Isolation System was used to isolated RNA from the syncytiotrophoblast of human term placentas. The isolated RNA was used for RT-PCR with M-MLV Reverse Transcriptase used for the RT reaction. The 110 kDa PKD2 protein was expressed in vitro with the TNT^® T7 Coupled Reticulocyte Lysate System with and without Canine Pancreatic Microsomal Membranes. The channel activity of the expressed protein was examined and the functional channel could be inhibited by known inhibitors. Expressing the luciferase control with the membranes did not produce the same effect. (2187)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from primary human coronary artery endothelial cells. The isolated RNA was used to make gene array ³³P-labeled targets on nylon cDNA microarray filters using Oligo(dT) and M-MLV Reverse Transcriptase, RNase H-. The same material was used for first-strand synthesis in RT-PCR. (2697)

Notes: The SV Total RNA Isolation System was used to isolated RNA from autopsy samples of human brain with FTDP-17 disorder. The isolated RNA was used for semi-quantitative RT-PCR (2162)

Notes: The study of a carbapenem resistant strain of Klebsiella pneumoniae lead to the description of a novel form of β-lactamase, which confers resistance to imipenem, meropenem, extended-spectrum cepahlosporins, and aztreonam. Total RNA used in a primer extension assay was isolated with the SV Total RNA Isolation System . Primer extension reactions were performed with Promega's Primer Extension System- AMV Reverse Transcriptase to determine the transcriptional start site of the KPC-1 β-lactamase gene. The KPC-1 β-lactamase gene was then sequenced with the fmol® DNA Sequencing System (2300)

Notes: Total RNA was isolated from Mycobacterium smegmatis for Northern blot analysis and RT-PCR. Transcripts for furA and katG were amplified by RT-PCR using Promega's MMLV Reverse Transcriptase and cloned into the pGEM^®-T Easy vector. Northern blot probes for furA and katG were synthesized by in vitro transcription from the pGEM^®-T Easy vector. (2310)

Notes: Total RNA was isolated from Rhodopseudomonas palustris using the SV Total RNA Isolation System. The Access RT-PCR System was used to determine the transcriptional organization of the badHIaliBA and badK genes, genes involved in anaerobic benzoate degradation. (2306)

Notes: Total RNA was isolated from Alteromonas sp. strain O-7 for Northern blot analysis of the aprII gene to show that aprII is upregulated under iron rich growth conditions. The transcriptional start site of this gene was mapped in a primer extension reaction using Promega's AMV reverse transcriptase. (2304)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from cultured human fibroblasts. The isolated RNA was used for RT-PCR. (2184)

Notes: The SV Total RNA Isolation System was used to isolate RNA from transiently transfected COS-7 cells bearing expression plasmid for exon 6 from a variety of patients. The isolated RNA was used for RT-PCR analysis. (2185)

Notes: The authors described a cluster of 8 genes from P. putida that is thought to be involved in benzoate metabolism: benA, benB, benC, benD, benE, benF, benK, and benR. The transcriptional start site of benA was determined using Promega's Primer Extension System-AMV Reverse Transcriptase. Total RNA used in these primer extension reactions was isolated from P. putida cells using the SV Total RNA Isolation System. The transcriptional organization of the benA, benB, and benC genes was determined by RT-PCR using the Access RT-PCR System. (2301)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from rat endothelial cells. The isolated RNA was used for RT-PCR and semi-quantitative RT-PCR with cDNA generated with AMV Reverse Transcriptase. (2175)

Notes: Jurkat cells were fractionated into polysomes, and the resulting polysomes were ethanol precipitated. Total RNA was isolated from the polysomes with the SV Total RNA Isolation System. The system was also used to isolate RNA directly from Jurkat cells without fractionation. The isolated RNA was used for Northern blots, dot blots, RT-PCR and RNase protection assays. (2182)