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Forensic Sci. Int. 138, 116–118. Population data for 11 STR loci in northeast China Han. 2003

Zhanga, Y.J., Xub, Q.S. and Lee, J.B.

Notes: In this paper, researchers report allele frequencies for a population in Northeastern China. Data was generated using the PowerPlex® 16 System. DNA from volunteers was extracted using a phenol chloroform method. (3000)

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Forensic Sci. Int. 138, 116–8. Population data for 11 STR loci in northeast China Han. 2003

Zhang, Y.J., Xu, Q.S. and Lee, J.B.

Notes: Allele frequencies for 11 autosomal STR loci were determined in the Yan Bian region of China. Population data for nine STR loci were determined using the AmpFlSTR® Profiler Plus™ kit and for the Penta D and Penta E loci using the PowerPlex® 16 System. DNA was isolated using phenol:chloroform extraction, and 1ng was amplified as directed by the manufacturer. Amplified products were detected using an ABI PRISM® 377 DNA Sequencer. (3826)

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Int. Congr. Ser. 1239, 235–8. Portuguese population data on two pentanucleotide STR loci Penta E and Penta D. 2003

Ribeiro, T., Viriato, L., Vieira-Silva, C., Cruz, C., Espinheira, R. and Geada, H.

Notes: The authors reported population data for the Penta E and Penta D loci in 160 Portuguese Caucasians, 19 Portuguese of African origin and 150 meioses. DNA was extracted from blood samples using Chelex® resin, then purified using the Wizard® DNA Clean-Up System. Amplifications were performed using the PowerPlex® 16 System, the SGM Plus® kit and the GeneAmp® PCR System 9600. Amplified products were detected using an ABI PRISM® 377 DNA Sequencer. (3845)

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Int. Congr. Ser. 1239, 125–30. PowerPlex™ 16 analysis in the Japanese population. 2003

Hashiyada, M., Itakura, Y. and Nata, M.

Notes: The authors generated population data from 508 unrelated Japanese individuals using the PowerPlex® 16 System. DNA was collected as buccal swabs and isolated using Chelex® resin followed by phenol:chloroform extraction and ethanol precipitation. For each PowerPlex® 16 reaction, 2ng of DNA was amplified using a GeneAmp® PCR System 9700, and amplified products were detected using the ABI PRISM® 377 DNA Sequencer. (3843)

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Clin. Diagn. Lab. Immunol. 10, 339 - 344. Production of genetically engineered biotinylated interleukin-2 and its application in a rapid nonradioactive assay for T-cell activation. 2003

Jordan, R.A., Preissler, M.T., Banas, J.A. and Gosselin, E.J.

Notes: Researchers used PCR primers with Kpn I and Hind III restriction sites to clone human IL-2 from a known rhIL-2 E. coli clone containing the PTCGF-11 vector. The PCR product was digested with Kpn I and Hind III and cloned into the PinPoint™ Xa-3 Vector. Transformed E. coli JM109 clones were then pre-incubated in the presence of 8μM biotin for 2 hours before being induced with 100μM IPTG for an additional 2 hours. After induction, the cells were collected and resuspended in a lysis buffer before mechanical lysis with a French press. The lysate was then passed over a SoftLink™ Soft Release Avidin Resin column and the biotinylated rhIL-2 eluted. The resultant purified biotinylated rhIL-2 displayed similar properties and biological activity to native IL-2. Elutants from cells transformed with the PinPoint™ Xa Control Vector produced no biotinylated rhIL-2 and did not display any properties indicating that IL-2 was present. (2680)

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Biol. Reprod. 69, 1060–1068. Production of nuclear transfer-derived piglets using porcine fetal fibroblasts transfected with the enhanced green fluorescent protein. 2003

Hyun, S., Lee, G., Kim, D., Kim, .H, Lee, S., Nam, D., Jeong, Y., Kim, S., Yeom, S., Kang, S., Han, J., Lee, B. and Hwang, W.

Notes: The authors of this report developed a means of somatic cell nuclear transfer (SCNT) that led to the successful production of GFP-transfected piglets. Citing the high value of transgenic pigs in biotechnology applications, the authors conducted these studies to establish a system for the production of transgenic pigs using SCNT of GFP-transfected cells into enucleated oocytes. FuGENE® 6 Transfection Reagent was used to transfect enhanced-GFP into confluent fetal pig fibroblasts (1µg of pEGFP-N1 with 3µl of FuGENE® 6 reagent in a 35mm dish). The cells were cultured for 2–3 days until confluency and passaged once to achieve stable integration of the gene into chromosomes before use for SCNT. (4281)

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J. Cell Sci. 116, 2421-2430. PTHrP [67–86] regulates the expression of stress proteins in breast cancer cells inducing modifications in urokinase-plasminogen activator and MMP-1 expression. 2003

Luparello, C., Sirchia, R. and Pupello, D.

Notes: Total and poly(A)+ RNA from treated and untreated 8701-BC breast cancer tumor cells were treated with RQ1 RNase-free DNase. The RNA samples were reverse transcribed using the M-MLV RNase H– point mutant reverse transcriptase and random primers to create cDNA used in downstream analyses. The cDNA from the reactions were used in PCR, differential display PCR, and semi-quantitative multiplex PCR reactions. In differential display PCR studies, the authors analyzed differentially displayed bands by staining 6% polyacrylamide gels with the SILVER SEQUENCE™ Staining Reagents. Differentially displayed bands were re-amplified and cloned using the pGEM®-T Easy Vector and high efficiency JM109 competent cells. (3020)

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Anal. Biochem. 316, 259–269. Quantitative intra-short interspersed element PCR for species-specific DNA identification. 2003

Walker, J.A., Hughes, D.A., Anders, B.A., Shewale, J., Sinha, S.K. and Batzer, M.A.

Notes: Cow (Bos taurus), horse (Equus caballus), sheep (Ovis aries), antelope (Antilocapra americana), dog (Canis familiaris), cat (Felis catus), and rabbit (Oryctolagus cuniculus) genomic DNA were obtained by extraction from tissue and blood samples using the Wizard® Genomic DNA Purification kit. SINEs (short interspersed elements) PCR was performed on various combinations of purified DNA from the above species. (3260)

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Biochem. J. 374, 359–367. Regulation of alternative splicing of Gtf2ird1and its impact on slow muscle promoter activity. 2003

Tay, E.S.E., Guven, K.L., Subramaniam, N., Polly, P., Issa, L.L., Hardeman, P.W. and Hardeman, E.C.

Notes: In this paper, ImProm-II™ Reverse Transcriptase was used to make cDNAs of MusTRD using template RNA isolated from C2C12 myotubes (C2C12) and various mouse hind limb muscle tissues. One microliter of the cDNA products from the reaction was used in real-time PCR analysis using primers for the alternately spliced MusTRD products and SYBR® Green Dye.  Data is presented as relative expression compared with the C2C12 cell line or as gel images of the resultant amplimers.  (2844)

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FEBS Lett. 554, 119-126. Regulation of the ABA-sensitive Arabidopsis potassium channel gene GORK in response to water stress. 2003

Becker, D., Hoth, S., Ache, P., Wenkel, S., Roelfsema, M.R.G., Meyerho, O., Hartung, W. and Hedrich, R.

Notes: M-MLV Reverse Transcriptase, RNase H Minus, was used to make first-strand cDNA from total and mRNA samples isolated from Arabidopsis seedlings and cell culture. cDNA was then diluted 1:20 in real-time PCR to quantitatively assess GORK, PP2CA, AKT2/3, KIN2 transcript levels. Data was normalized to actin transcript levels.  (3022)

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Am. J. Physiol. Renal Physiol. 284, F829–F839. Renal tubular epithelial cell apoptosis is associated with caspase cleavage of the NHE1 Na+/H+ exchanger. 2003

Wu, K.L., Khan, S., Lakhe-Reddy, S., Wang, L., Jarad, G., Miller, R.T., Konieczkowski, M., Brown, A.M., Sedor, J.R. and Schelling, J.R.

Notes: This paper describes amplification of rat NHE1 cDNA using primers incorporating the Xho I and Xba I restriction endonuclease sites, Kozak consensus, ATG start site, and a stop codon. The resultant 1.1 kb NHE1 encoding-PCR product was digested with Xba I and Xho I and cloned into the pTNT™ Vector.  This construct was used as a template in a TNT® T7 Quick Coupled Transcription/Translation reaction to generate substrates for an in vitro caspase-3 cleavage assay. Proteolytically cleaved fragments of NHE1 were run on a polyacrylamide gel and visualized by autoradiography. (3063)

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J. Biol. Chem. 278 (35), 33384–33391. Role of the glucocorticoid receptor for regulation of hypoxia-dependent gene expression. 2003

Kodama, T., Shimizu, N., Yoshikawa, N., Makino, Y., Ouchida, R., Okamoto, K., Hisada, T., Nakamura, H., Morimoto, C. and Tanaka, H.

Notes: The SV Total RNA Isolation System was used to isolate total RNA from HeLa cells for RT-PCR analysis. Fifty nanograms of the total RNA was used as template in AccessQuick™ RT-PCR System reactions. VEGF, ADM, GLUT3, and β-actin mRNA levels in HeLa cells under normoxia and hypoxia conditions were analyzed. (2746)

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Plant J. 36(3), 421-429. Sequence database of 1172 T-DNA insertion sites in Arabidopsis activation-tagging lines that showed phenotypes in T1 generation. 2003

Ichikawa, T., Nakazawa, M., Kawashima, M., Muto, S., Gohda, K., Suzuki, K., Ishikawa, A., Kobayashi, H., Yoshizumi, T., Tsumoto, Y., Tsuhara, Y., Iizumi, H., Goto, Y. and Matsui, M.

Notes: To find gain-of-function mutants in Arabidopsis, T-DNA insertions were screened for visible phenotypes. 1172 plants were found to have gain-of-function insertions. To map the insertion site of the T-DNA, genomic DNA was isolated on an automated workstation (Tecan Genesis®) using the Wizard® Magnetic 96 DNA Plant System The original plasmid was then isolated from the genomic DNA after digestion and transformation into bacteria. The plasmid was sequenced to determine the site of insertion. (2788)

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Proc. Natl. Acad. Sci. USA 100, 13773-13778. Shade is the Drosophila P450 enzyme that mediates the hydroxylation of ecdysone to the steroid insect molting hormone 20-hydroxyecdysone. 2003

Petryk, A., Warren, J.T., Marqués, G., Jarcho, M.P., Gilbert, L.I., Kahler, J., Parvy, J.P., Li, Y., Dauphin-Villemant, C. and O'Connor, M.B.,

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)

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Mol. Pharmacol. 63(4), 915-924. Species-specific transcriptional activity of synthetic flavonoids in guinea pig and mouse cells as a result of differential activation of the aryl hydrocarbon receptor to interact with dioxin-responsive elements 2003

Zhou, J.G., Henry, E.C., Palermo, C.M., Dertinger, S.D. and Gasiewicz, T.A.

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)

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J. Cell Sci. 115, 1643-1649. Stem cell factor activates telomerase in mouse mitotic spermatogonia and in primordial germ cells. 2003

Dolci, S., Levati, L., Pellegrini, M., Faraoni, I., Graziani, G., Di Carlo, A. and Geremia, R.

Notes: RT-PCR was used to assess the response of mitotic spermatogonia to Kit1. Spermatogonia were cultured with and without Kit1 and RNA harvested after 24 hours. RT-PCR was performed in a two-step reaction using first the ImProm-II™ Reverse Transcriptase in a 20µl reaction. One microliter of the RT reaction was removed and amplified in a 50µl PCR using the PCR Master Mix. (2626)

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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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