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J. Neurosci. 23, 5012–5019. Stoichiometry of expressed KCNQ2/KCNQ3 potassium channels and subunit composition of native ganglionic M channels deduced from block by tetraethylammonium. 2003

Hadley, J.K., Passmore, G.M., Tatulian, L., Al-Qatari, M., Ye, F., Wickenden, A.D. and Brown, D.A.

Notes: Single cell RT-PCR was performed using M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant, and the extracted cytosol of sympathetic neurons isolated from 17-day-old or 45-day-old rats. An oligo(dT) primer was used in the reverse transcription reactions. The resultant cDNA was used in PCR with KCNQ2-, KCNQ3-, KCNQ4-, and KCNQ5-specific, potassium channel subunit gene primers.  (3024)

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J. Immunol. 171(5), 2270-8. Systemic overexpression of IL-10 induces CD4+ CD25+ cell populations in vivo and ameliorates type 1 diabetes in nonobese diabetic mice in a dose-dependent fashion. 2003

Goudy, K.S., Burkhardt, B.R., Wasserfall, C., Song, S., Campbell-Thompson, M.L., Brusko, T., Powers, M.A., Clare-Salzler, M.J., Sobel, E.S., Ellis, T.M., Flotte, T.R. and Atkinson, M.A.

Notes: Nonobese diabetic (NOD) mice were infected with a nonpathogenic recombinant adeno-associated virus carrying the IL-10 gene. To verify IL-10 transgene expression, total RNA was isolated and IL-10 specific primers were used for amplification using the AccessQuick™ RT-PCR System. (3093)

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Plant J. 34(6), 856-67. The Arabidopsis thaliana CUTA gene encodes an evolutionarily conserved copper binding chloroplast protein. 2003

Burkhead, J.L., Abdel-Ghany, S.E., Morrill, J.M., Pilon-Smits, E.A. and Pilon, M.

Notes: Messenger RNA was in vitro transcribed and capped using linearized plasmids containing the genes for preplastocyanin and  mature plastocyanin, a copper binding protein. Then, 0.2 µg of each plastocyanin RNA was added to Wheat Germ Extract for translation. Translated proteins were labeled with 35S-methionine. (3108)

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Clin. Can. Res. 9, 2981-2984. The RASSF1A tumor suppressor gene is commonly inactivated in adenocarcinoma of the uterine cervix 2003

Cohen, Y., Singer, G., Lavie, O., Dong, S.M., Beller, U., Sidransky, D.

Notes: The authors used real-time methylation-specific PCR to detect and quantitate the bisulfite-converted methylated version of the RASSF1A promoter region in uterine carcinoma samples. Bisulfite-treated DNA was purified using the Wizard® DNA Clean-Up System. (2710)

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Development 130, 5481 - 5491. The role of neurotrophin receptors in female germ-cell survival in mouse and human. 2003

Spears, N., Molinek, M.D., Robinson, L.L.L., Fulton, N., Cameron, H., Shimoda, K., Telfer, E.E., Anderson, R.A., and Price, D.J.

Notes: TrkB was shown to be important in the survival of germ cells during follicle formation in the ovaries. Immunohistochemical analysis of mouse ovaries showed the presence of TrkB. TrkB was found in similar patterns in germ cells both before and after follicle formation, but TrkB protein was more abundant in P0 oocytes than in E16 oogonia. IHC was performed on ovaries fixed in Bouin's fixative then embedded in wax. Anti-TrkB In pAb was used at a 1:10 dilution.   (2829)

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Appl. Environ. Microbiol. 69 (7), 3952–3956. Transfection of Diaporthe perjuncta with Diaporthe RNA Virus. 2003

Moleleki, N., van Heerden, S.W., Wingfield, M.J., Wingfield, B.D. and Preisig, O.

Notes: A full length cDNA representing the entire genome of Diaporthe RNA virus (DaRV) was successfully cloned into the pGEM®-T Easy Vector. The resultant construct was named pDV3. A positive strand viral RNA was then synthesized from the Sal I linearized pDV3 template. Fungi transfected viral RNA were shown to have different morphological features when compared to untransfected fungi. (2757)

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Clin. Can. Res. 9, 3190-3197. Tumor growth retardation, cure, and induction of antitumor immunity in B16 melanoma-bearing mice by low electric field-enchanced chemotherapy 2003

Entin, I., Plotnikov, A., Korenstein, R., Keisari, Y.

Notes: Reverse transcription and PCR was performed on total RNA isolated from spleens of treated mice to determine the effect of treatment on cytokine mRNA expression using the Reverse Transcription System. (2718)

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J. Forensic Sci. 47, 773-785. Validation of a 16-locus fluorescent multiplex system. 2003

Krenke, B.E., Tereba, A., Anderson, S.J., Buel, E., Culhane, S., Finis, C.J., Tomsey, C.S., Zachetti, J.M., Masibay, A., Rabbach, D.R., Amiott, E.A. and Sprecher, C.J.

Notes: In this paper, researchers from both Promega and state crime laboratories provide an evaluation of the Powerplex® 16 System loci (CODIS, Penta D, Penta E, and amelogenin). Data from several laboratories using a variety of thermal cyclers and the ABI PRISM® 310 Genetic Analyzer or ABI PRISM® 377 DNA Sequencer are presented. The Powerplex® 16 System was able to consistently genotype samples from as little as 0.0625-2ng of genomic DNA template. Other factors such as reaction volume, annealing temperature, titration of AmpliTaq Gold® DNA Polymerase, magnesium, or primer pairs, and 1ng DNA mixtures were analyzed using the Powerplex® 16 System.  The researchers also present data on stutter analysis and cross reactivity of primer sets on primate DNA templates for each loci. (2675)

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J. Biol. Chem. 278, 5156-5162. Y-box-binding protein YB-1 mediates transcriptional repression of human alpha2(I) collagen gene expression by interferon-gamma. 2003

Higashi, K., Inagaki, Y., Suzuki, N., Mitsui, S., Mauviel, A., Kaneko, H. and Nakatsuka, I.

Notes: The Y-box-binding protein YB-1 was examined through the use of a reporter plasmid with wildtype and mutant putative Y-box binding sites. The binding sites were constructed in the pGL3 Basic Vector and measured using the Luciferase Assay System. Studies were performed in normal human dermal fibroblasts.  Expression of YB-1 was found to repress expression of the COL1A2 gene at the transcriptional level. Confirmation of this dose-dependent inhibition was through measurement of the steady-state mRNA levels by quantitative, real-time RT-PCR.  The reverse transcription portion of the quantitative, real-time RT-PCR reactions were performed with ImProm-II™ Reverse Transcriptase followed by a TaqMan-type quantitative PCR step. (2623)

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Clin. Can. Res. 8, 1800-1807. A novel gene from the human endogenous retrovirus K expressed in transformed cells 2002

Armbruester, V., Sauter, M., Krautkraemer, Meese, E., Kleiman, A., Best, B., Roemer, K., and Mueller-Lantzsch, N.

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)

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Endocrinology 142, 4320-4330. Aldosterone increases T-type calcium currents in human adrenocarcinoma (H295R) cells by inducing channel expression. 2002

Lesouhaitier, O., Chiappe, A., and Rossier, M.F.

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)

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Mol. Cell. Neurosci. 19, 447-458. AMPA Receptor-Mediated Modulation of Inward Rectifier K+ Channels in Astrocytes of Mouse Hippocampus 2002

Schroder, W., Seifert, G., Huttman, K., Hinterkeuser, S., Steinhauser, C.

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)

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J. Virol. 76, 12032-12043. Baculorus lef-12 is not required for viral replication. 2002

Guarino, L.A. Mistretta, T.-A. and Dong, W.

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)

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J. Forensic Sci. 47(4), 757-772. Characterization and validation studies of PowerPlex® 2.1, a nine-locus short tandem repeat (STR) multiplex system and Penta D monoplex. 2002

Levedakou, E.N., Freeman, D.A, Budzynski, M.J., Early, B.E., Damaso, R.C., Pollard, A.M., Townley, A.J., Gombos, J.L., Lewis, J.L., Kist, F.G., Hockensmith, M.E., Terwilliger, M.L., Amiott, E., McElfresh, K.C., Schumm, J.W., Ulery, S.R., Konotop, F., Sessa, T.L., Sailus, J.S., Crouse, C.A., Tomsey, C.S., Ban, J.D. and Nelson, M.S.

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)

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Infect. Immun. 70, 4389–4398. Characterization of Pit, a Streptococcus pneumoniae iron uptake ABC transporter. 2002

Brown, J.S., Gilliland, S.M., Ruiz-Albert, J. and Holden, D.W.

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)

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Hum. Mol. Genet. 11, 2701–2708. Cholesterol regulates ABCD2 expression: Implications for the therapy of X-linked adrenoleukodystrophy.
2002

Weinhofer, I., Forss-Petter, S., Zigman, M. and Berger, J.

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)

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Genetics 160, 225-234. Cloning and characterization of the Tribolium cataneum eye-color genes encoding tryptophan oxygenase and kynurenine 3-monooxygenase 2002

Lorenzen, M.D., Brown, S.J., Dennell, R.E., and Beeman, R.W.

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)

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Invest. Ophthalmol. Vis. Sci. 43, 72-81. Differential effect of activin A and BMP-7 on myofibroblast differentiation and the role of the Smad signaling pathway. 2002

You, L. and Kruse, F.E.

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)

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Plant Physiol. 130, 58–67. Differential expression of a metallothionein gene during the presymbiotic versus the symbiotic phase of an arbuscular mycorrhizal fungus. 2002

Lanfranco, L., Bolchi, A., Ros, E.C., Ottonello, S. and Bonfante, P.

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)

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Biochem. J. 372, 347–357. Differential functional properties of calmodulin-dependent protein kinase IIã variants isolated from smooth muscle. 2002

Gangopadhyay, S.S., Barber, A.L., Gallant, C., Grabarek, Z., Smith J.L., and Morgan, K.G.

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)

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Cell Death Differ. 9, 1334-1342. Differential subcellular localization of functionally divergent survivin splice mutants 2002

Mahotka, C., Liebmann, J., Wenzel, M., Suschek, C.V., Schmitt, M., Gabbert, H.E., Gerharz, C.D.

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)

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Proc. Natl. Acad. Sci. USA 99, 54–59. Efficient bacterial transcription of DNA nanocircle vectors with optimized single-stranded promoters. 2002

Ohmichi, T., Maki, A. and Kool, E.T.

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)

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J. Biol. Chem. 277, 42280–42288. GAGA factor down-regulates its own promoter. 2002

Kosoy, A., Pagans, S., Espina´s, M.L., Azorýn, F. and Bernue´s, J.

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)

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Forensic Sci. Int. 128, 196–201. Genetic diversity at two pentanucleotide STR and thirteen tetranucleotide STR loci by multiplex PCR in four predominant population groups of central India. 2002

Sarkar, N. and Kashyap, V.K.

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)

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Infect. Immun. 70, 1984–1990. Helicobacter pylori uses motility for initial colonization and to attain robust infection. 2002

Ottemann K.M. and Lowenthal, A.C.

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)

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