Notes: The authors used the pGEM-T vector to clone a novel gene identified through RT-PCR analysis of human endogenous retrovirus K (HERV-K). (2469)

Notes: Total RNA was isolated from 1 x 10⁶ H295R cells using the RNAgents^® Total RNA Isolation System. The isolated RNA was used for conventional and real-time quantitative RT-PCR. (2565)

Notes: The patch-clamp technique was combined with RT-PCR in acute hippocampal brain slices. After recording, each cell was harvested to perform transcript analysis and identify subunits that underlie inwardly rectifying K+ currents. The recording pipette solution was supplemented with recombinant RNasin Ribonuclease Inhibitor. After recording channel activity, cell contents were harvested through the recording pipette. RT-PCR was performed on cell contents. (2426)

Notes: RT-PCR was used to confirm the expression of the lef-12 gene at various times post-infection. The RT step was performed using ImProm-II™ Reverse Transcriptase in a 20µl reaction volume. One microliter of the RT reaction was amplified in a 50µl PCR amplification reaction using the PCR Master Mix. (2628)

Notes: This study provides a multi-lab validation of the PowerPlex^® 2.1 and Penta D systems. The paper also reveals the primer sequences for all ten loci. Stutter, species cross-reactivity, environmental variations, and sample mixtures were examined for all loci. The researchers were able to consistently amplify from 0.75ng of sample DNA.  (2679)

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: The sterol regulatory element-binding protein-1a (SREBP1a) was in vitro transcribed and translated from a plasmid template using TNT^® T7 Quick for PCR DNA. A control reaction was performed using Transcend™ tRNA to confirm that the 120KDa protein was expressed correctly.  Unlabeled SREBP1a was used in gel-shift assays with a labeled oligo representing the SRE motif from the human ABCD2 gene promoter.  (3222)

Notes: The Wizard^® Genomic DNA Purification Kit was used to isolate genomic DNA from individual Tribolium cataneum flour beetles. The isolated genomic DNA was used for PCR amplification for recombinational mapping related to eye color, PCR-based deletion breakpoint analysis, and was EcoR I-digested for Southern blotting analysis. (2504)

Notes: Total RNA was isolated from human corneal tissue with the RNAgents^® Total RNA Isolation System.  The total RNA was further processed into the poly(A)+ fraction through the use of the PolyATtract^® mRNA Isolation System. The isolated mRNA was used for RT-PCR and Northern analysis. RT-PCR was performed using M-MLV Reverse Transcriptase. (2569)

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: Promega reverse transcription buffer, AMV reverse transcriptase, and RNAsin^® RNAse inhibitor were used in reverse transcription reactions of total RNA from HepG2 heptatoma cells. PCR products were cloned into the pGEMT vector. (2613)

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: Researchers PCR amplified and cloned the Trl promoter into the pGEM^®-T vector. The Trl promoter sequence, as well as the eve promoter sequence and/or a CMV promoter, were used to make luciferase reporter constructs using the pGL3-Basic Vector. Transient transfection experiments were performed with the promoter constructs and GAGA factor expression constructs. Relative luciferase activity from each experiment was measured and compared to that of a control.  (2775)

Notes: Population data was determined for 208 unrelated individuals from central India using the PowerPlex^® 16 System. DNA was isolated from blood by organic extraction, and 2ng was amplified per reaction. Amplified products were detected using an ABI PRISM^® 377 DNA Sequencer. (3829)

Notes: The authors of this paper describe using the Wizard^® Genomic DNA Purification Kit to isolate genomic DNA from Helicobacter pylori. The isolated genomic DNA was used in PCR to analyze motB gene mutations.  (3101)

Notes: Genomic DNA was isolated from H. pylori with the Wizard® Genomic DNA Purification Kit. The isolated DNA was used in both PCR amplification and Southern blotting. (2292)

Notes: In this paper, biotinylated and fluorescently labeled DNA probes were used simultaneously in hybridization reactions with restriction digested human genomic DNA. Streptavidin-paramagnetic particles were used to capture restriction fragments of interest prior to analysis for CpG methylation and short-nucleotide repeats with an alkaline-phosphatase-conjugated anti-fluorescein antibody.  These immuno-DNA complexes were then analyzed using the AttoPhos^® AP Fluorescent Substrate System in 96-well plates. Fluorescence signal was determined using a FluoroCount™ microplate reader. Details of  hybridization conditions, probe concentrations, washes and statistical analysis are provided. (2655)

Notes: The pGEM^®-T Easy Vector was used to clone a 12.5 kb fragment containing human kidney specific chloride channel (CLC-KB) gene fused to an enhanced green fluorescence protein (EGFP) created in a long and accurate PCR (LA-PCR) reaction.  The resultant construct was further manipulated and used to make transgenic mice.  (3131)

Notes: This study investigated the effect of aberrant expression of EAAT2 mRNA on expression of the EAAT2 protein in human glioma cells. EAAT2 encodes an excitatory amino acid transporter that helps to clear glutamate from synaptic cleft. In transfection-based studies on human U251 gliomal cells,  transient transfection efficiency was evaluated using a luciferase reporter gene, and luciferase activity was detected with the Luciferase Assay System. In separate experiments, RTPCR products were cloned into plasmid vectors. Plasmid DNAs containing PCR products were isolated from bacteria using the Wizard^® Plus SV Minipreps DNA Purification System. (2607)

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: The Wizard^® Genomic DNA Purification Kit was used to purify DNA from two strains of Aspergillus fumigatus.  A modified protocol was used for the purification.  Cells were filtered through a 0.2μm SFCA filter, frozen on dry ice and then ground with a mortar and pestle.  The ground cells were then incubated  in Nuclei Lysis Solution at 37°C for 1 h before continuing with the Wizard^® Genomic DNA Tissue Culture Cell procedure.  The purified DNA was used in PCR with primers for the A. fumigatus PMA1 gene. (3048)

Notes: Total RNA was isolated from various mouse tissues including brain and cultured cells using the RNAgents^® Total RNA Isolation System. The isolated RNA was used for both RACE and RT-PCR analysis. (2579)

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: In this paper, total RNA isolated from transgenic mouse pre-B cells was used as template for semi-quantitative RT-PCR. The RNA was isolated using the RNAgents^® Total RNA Isolation System. Promoter studies were performed in 293T cells with the Dual-Luciferase^® Reporter Assay System. Firefly luciferase activity controlled by Renilla luciferase activity was provided by the pRL-TK Vector. (2566)