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Mol. Cell. Biol. 25, 2861-2870. Assembly of the mariner Mos1 synaptic complex. 2005

Auge-Gouillou, C., Brillet, B., Hamelin, M.H., and Bigot, Y.

Notes: A transposase protein (Tnp) open reading frame (amino acids 1 to 345) was amplified from the Drosophila mauritiana Mos1 mariner transposable element using GoTaq® DNA polymerase. The Tnp open reading frame was then cloned with into the pGEM®-T Easy Vector and sequenced before further subloning. Tnp protein was purified and used in gel-shift assays to demonstrate Tnp binding regions in the Mos1 transposable element. (3344)

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Int. J. Radiat. Oncol. Biol. Phys. 61, 517-28. Bcl-2 inhibitors potentiate the cytotoxic effects of radiation in Bcl-2 overexpressing radioresistant tumor cells. 2005

Hara, T., Omura-Minamisawa, M., Chao, C., Nakagami, Y., Ito, M. and Inque, T.

Notes: Total RNA was isolated from HeLa cells (wild-type and stably transformed) and 2μg of the total RNA was reverse transcribed into cDNA. PCR was performed using 1 or 2μl of the cDNA, 0.5μM of each gene-specific primer (GAPDH, Bcl-2, or Bcl-xl), 0.2mM dNTP mix and GoTaq® DNA Polymerase (5U/μl) with the corresponding 5X GoTaq® Reaction Buffer in a final volume of 20μl. The initial denaturation step at 95°C for 2 minutes was followed by 25 cycles of denaturation at 94°C for 30 seconds (15 seconds for GAPDH), annealing at 55°C for 30 seconds, extension at 72°C for 30 seconds, and a final extension step at 72°C for 10 minutes. The PCR products were separated on 2% agarose gels. (3369)

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Plant Physiol. 137(2), 557–66. Beyond complementation. Map-based cloning in Chlamydomonas reinhardtii. 2005

Rymarquis, L.A., Handley, J.M., Thomas, M. and Stern, D.B.

Notes: These authors describe map-based cloning tools for Chlamydomonas reinhardtii, (green yeast). In this case-based study of existing and projected resources for C. reinhardtii mapping, mcd4 and mcd5 mutants were mapped by crossing to the infertile strain known as Chlamydomonas grossii, S1-D2, with several known marker sites. To reduce the time and expense of mapping, bulked segregant analysis (BSA) and marker duplexing were evaluated. In BSA, DNAs from multiple segregating progeny (up to six in this study) are combined, and results from PCR-based markers are examined for significant bias from a roughly equal contribution from each parent. This case study used 57–72 markers to span the 1,107-cM genome using approximately 3,350 PCR amplifications. The researchers picked single colonies from a plate and placed each colony in a hypotonic EDTA solution. After boiling for 5 minutes, the supernatant was collected by centrifugation and the DNA quantitated. Twenty nanograms of DNA thus isolated was used as the template for PCR. The reaction included GoTaq® DNA polymerase with the enhancers 8.5%glycerol and 0.83% formamide in a 30µl reaction. Forty cycles of amplification were performed and the results were analyzed by agarose gel electrophoresis. The various amplification reactions included the use of BSA and both monoplex and duplex reactions with amplicons of 90bp–625bp. (3279)

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J. Hospital Infect. 61, 225-230. Detection of icaA and icaD loci by polymerase chain reaction and biofilm formation by Staphylococcus epidermidis isolated from dialysate and needles in a dialysis unit. 2005

Chaieb, K., Mahdouani, K., and Bakhroug, A.

Notes: The presence or absence of intercellular adhesion genes (icaA and icaD) in various strains of Staphylococcus epidermidis isolated in medical facilities was investigated using PCR amplification and GoTaq® DNA Polymerase. Each reaction contained 150ng of template. GoTaq® Green Reaction Buffer was used in the PCR. (3359)

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J. Eukaryot. Microbiol. 52, 31-37. Development and molecular characterisation of the microsporidian Schroedera airthreyi n. sp. in a freshwater bryozoan Plumatella sp. (Bryozoa: Phylactolaemata). 2005

Morris, D.J., Terry, R.S., and Adams, A.

Notes: GoTaq® DNA Polymerase was used to amplify various portions of the rDNA isolated from microsporidia-infected brysozoans. The sequence of these amplimers was used in the construction of a phylogenetic tree. (3379)

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FEBS Lett. 279, 2514-8. Gene expression and characterization of isoprene synthase from Populus alba 2005

Sasaki, K., Ohara, K. and Yazaki, K.

Notes: In vitro cultures of P. alba were incubated in growth chambers under various conditions. The axenic plants of P. alba were sprayed with either methyl jasmonate (100 lM), methyl salicylate (1mM), or ethanol as a control, and then grown under illumination at 18 hours/day for 24 hours at 25°C. Total RNA was subjected to semi-quantitative RT-PCR using GoTaq® DNA polymerase and primers for hybrid poplar isoprene synthase. For normalization, actin was used as an external standard. (3355)

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Mol. Cell. Biol. 25, 5763-76. Generation and characterization of Rac3 knockout mice. 2005

Corbetta, S., Gualdoni, S., Albertinazzi, C., Paris, S., Croci, L., Giacomo Consalez, G., and de Curtis, I.

Notes: GoTaq® DNA Polymerase was used in an RT-PCR. Aliquots of RNA were transcribed into cDNA and amplified with GoTaq® DNA Polymerase using primers specific for Rac1. (3351)

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Mol. Cell. Biol. 25, 5763–76. Generation and characterization of Rac3 knockout mice. 2005

Corbetta, S., Gualdoni, S., Albertinazzi, C., Paris, S., Croci, L., Consalez, G.G. and de Curtis, I.

Notes: To examine the role of Rac3 in nervous system development, knockout mice were created. Total RNA from postnatal day 0 (P0) and day 9 (P9) brains from wild-type (+/+), heterozygous (+/-), and knockout (-/-) mice was reverse transcribed and a ~260bp product amplified by PCR using GoTaq® DNA Polymerase and primers for either Rac3 or Rac1 (a Rac3 homolog). No amplification product for Rac3 was observed in the knockout mice. (3328)

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Vet. Parasitol. 133, 283-287. Genotyping of Giardia intestinalis from domestic and wild animals in Japan using glutamete dehydrogenase gene sequencing. 2005

Itagaki, T., Kinoshita, S., Aoki, M., Itoh, M., Saeki, H., Sato, N., Uetsuki, J., Izumiyama, S., Yagita, K., and Endo, T.

Notes: GoTaq® DNA Polymerase was used in PCR genotyping of Giardia intestinalis. Primers were designed around a 177 bp sequence of the glutamete dehydrogenase gene (gdh). Typing was based on previously reported assemblages of gdh from cats, dogs, cows and monkeys. PCR was performed in a total reaction volume of 25μl using 1.25 units of GoTaq® DNA Polymerase.
(3364)

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Intl. J. Parasitol. 35, 1071-1078. Molecular data suggest that microsporidian parasites in freshwater snails are diverse. 2005

McClymont, H.E., Dunn, A.M., Terry, R.S., Rollinson, D., Littlewood, D.T., and Smith, J.E.

Notes: Researchers tested several snail samples from various species for the presence of microsporidia infection. PCR tests were performed with GoTaq® DNA Polymerase and primers specific to microsporidia 16S DNA sequences. Isolated snail/microsporidia DNA was amplified in 25µl reactions with 0.625 units of GoTaq® DNA Polymerase. (3378)

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Infect. Immun. 73, 7064-7068. Mycobacterial lipomannan induces matrix metalloproteinase-9 expression in human macrophagic cells through a Toll-like receptor 1 (TLR1)/TLR2- and CD14-dependent mechanism. 2005

Elass, E., Aubry, L., Masson, M., Denys, A., Guérardel, Y., Maes, E., Legrand, D., Mazurier, J., and Kremer, L.

Notes: M-MLV Reverse Transcriptase was used to make cDNA from differentiated THP-1 cell total RNA. For these reverse transcription experiments the authors used 5 µg of total RNA, oligo dT and RNasin®. GoTaq® DNA Polymerase was used to amplify the cDNA from the reverse transcription reactions. PCR products were separated by gel electrophoresis and analyzed by ethidium bromide staining and band densitometry. (3357)

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RNA 11, 1571-8. Rat1p and Rai1p function with nuclear exosome in the processing and degradation of rRNA precursors. 2005

Fang, F., Phillips, S. and Butler, J.S.

Notes: GoTaq® DNA Polymerase was used in a RT-PCR. The 50μl PCR reactions contained 0.25μl cDNA, 12.5pmol primer, 0.025mM dNTPs, 2.5mM MgCl2 and 0.25μl of GoTaq® DNA Polymerase (5U/μl). PCR was performed using the following conditions: 94°C for 3 minutes followed by 20 cycles (94°C for 30 seconds, 55°C for 30 seconds and 72°C for 30 seconds), followed by 72°C for 7 minutes and held at 4°C. Products were visualized on an agarose gel. (3352)

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RNA 11, 1571–8. Rat1p and Rai1p function with the nuclear exosome in the processing and degradation of rRNA precursors. 2005

Fang, F., Phillips, S. and Butler, J.S.

Notes: To examine the effect of the exoribonuclease Rat1p on processing of ribosomal RNAs (rRNA), total RNA was isolated from several yeast strains with altered RAT1 expression. One microgram of DNase-treated total RNA was reverse transcribed in a 20µl reaction. Subsequently, 0.25µl of cDNA product was amplified using 0.25µl of GoTaq® DNA polymerase in a total volume of 50µl. The RT-PCR products were separated on 2% agarose gels and stained using ethidium bromide. (3330)

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J. Biol. Chem. 280, 36802–8. Structural and genetic analyses reveal a key role in prophage excision for the TorI response regulator inhibitor. 2005

Elantak, L., Ansaldi, M., Guerlesquin, F., Mejean, V. and Morelli, X.

Notes: To test the activity of torI as a prophage excisionase, the chloramphenical aceytltransferase gene was introduced into a non-coding region of the prophage KplE1 in a strain of E. coli containing a torI-encoding plasmid. Expression of torI was induced by IPTG and the cells plated on either chloramphenical or ampicillin. GoTaq® DNA polymerase was used in a PCR test to confirm prophage DNA excision from randomly chosen ampicillin-resistant colonies. (3329)

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J. Biol. Chem. 280, 36802-36808. Structural and genetic analyses reveal a key role in prophage excision for the torl response regulator inhibitor. 2005

ElAntak, L., Ansaldi, M., Guerlesquin, F., Méjean, V. and Morelli, X.

Notes: GoTaq® Polymerase was used in this study investigating the role of TorI (Tor inhibition protein) in prophage DNA excision. PCR products were amplified from E. coli strains carrying a plasmid-encoded torI gene, and were analyzed by gel electrophoresis. (3350)

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Nucl. Acids Res. 33(4), e41. Targeted 'knockdown' of spliceosome function in mammalian cells. 2005

Matter, N. and Konig, H.

Notes: To examine why there are two splicing systems present in multicellular organisms, the authors used morpholino oligomers complementary to the branch-site recognition elements of U2 or U12 small nuclear RNA to specifically suppress the splicing function in EL4 and Jurkat cells. After transfection with the morpholino oligos, the cells were harvested, and total RNA was isolated followed by DNase-treatment. In a 20µl reverse transcription reaction, 0.8µg total RNA was reverse transcribed with 100ng oligo(dT)15 primer and 200 units of M-MLV Reverse Transcriptase, RNase H Minus. PCR was then performed using 5µl of the diluted (1:10) reverse transcriptase reaction with 2.5 units GoTaq® DNA polymerase in a 50µl reaction for 28–31 cycles. (3278)

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Nucl. Acids Res. 33, e41. Targeted ‘knockdown’ of spliceosome function in mammalian cells. 2005

Matter, N. and König, H.

Notes: PCR was performed in 50μl GoTaq® Reaction Buffer containing 5μl of diluted (1:10) reverse transcriptase reactions, 250mM each dNTP, 10 pmol primers and 2.5 U GoTaq® DNA Polymerase. A total of 28 cycles were performed for amplification of P120 minigene transcripts, and 31 cycles were performed for endogenous P120 and ADPRT transcripts. Each cycle consisted of 95°C for 1 min, 59°C (P120) or 58°C (ADPRT) for 1 min and 72°C for 1.5 min. (3353)

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Mol. Cell. Biol. 24 (24), 10766–10776. 5-Fluorouracil enhances exosome-dependent accumulation of polyadenylated rRNAs. 2004

Fang, F., Hoskins, J. and Butler, J.S.

Notes: GoTaq® DNA Polymerase was used to amplify cDNAs created from total RNA of various yeast strains. Primers specific to ACT1 mRNA and 27S rRNA were used to analyze transcription levels under various conditions.  (3226)

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Toxicol. Sci. 82, 250–258. ARNT2 is not required for TCDD developmental toxicity in Zebrafish. 2004

Prasch, A.L., Heideman, W. and Peterson, R.E.

Notes: GoTaq® DNA Polymerase was used in PCR amplifications to genotype zfarnt2 -/- zebrafish mutants with a specific set of primers. The zfarnt2 gene encodes a receptor that is important for mediating chemical toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in zebrafish. (3192)

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Neurosci. Lett. 368, 41-45. Association study of polymorphisms in the 5’ upstream region of human DISC1 gene with schizophrenia. 2004

Kockelkorn, T.T., Arai, M., Matsumoto, H., Fukuda, N., Yamada, K., Minabe, Y., Toyota, T., Ujike, H., Sora, I., Mori, N., Yoshikawa, T. and Itokawa, M.

Notes: In this study, GoTaq® DNA Polymerase was used to PCR amplify a region of the Disrupted-in-Schizophrenia-1 (DISC1) gene from patient samples. Specific primers were used in the amplification reactions to test for polymorphisms in the region. (3128)

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Biochem. Biophys. Res. Commun. 321(1), 259-265. Cloning of hOST-PTP: the only example of a protein-tyrosine-phosphatase the function of which has been lost between rodent and human 2004

Cousin, W., Courseaux, A., Ladoux, A., Dani, C., and Peraldi, P.

Notes: Researchers used GoTaq® DNA polymerase to amplify 139bp and 815bp regions of hOST-PTP cDNA for detection and probe synthesis. The full-length 4006bp cDNA was amplified with Pfu DNA Polymerase. Fragments were gel purified using the Wizard® SV Gel and PCR Clean-Up System prior to cloning into the pGEM®-T Easy Vector.  The Prime-a-Gene® Labeling System was used to make 32P-dCTP labeled probes, which were used to screen cDNA clones. (3111)

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Oncogene 23, 8920-30. Fibronectin and type IV collagen activate ERα AF-1 by c-Src pathway: effect on breast cancer cell motility. 2004

Sisci, D., Aquila, S., Middea, E., Gentile, M., Maggiolini, M., Mastroiami, F., Montanaro, D. and Ando, S.

Notes: MCF-7 cells were incubated for 24 hours in PRF-SFM and then detached and plated on uncoated dishes or dishes coated with 2μg/cm2 poly-L-Lysine (P-Lys) in PBS, 30μg/ml fibronectin (Fn) in PBS or 30μg/ml type IV Collagen (Col) in 10mM acetic acid. The cells plated on uncoated dishes were treated both with and without 10nM estradiol (E2). After 24 hours, total cellular RNA was extracted and reverse transcribed using M-MLV reverse transcriptase. Briefly, reverse transcription was performed on 1μg of total RNA in a final volume of 10μl by incubation at 37°C for 30 min with 200U of M-MLV reverse transcriptase, 0.4μg oligo-dT, 0.5μM dNTP and 24U RNasin® Ribonuclease Inhibitor, followed by heat denaturation for 5 minutes at 95°C. Subsequent PCR analysis was performed on 1μl of the RT product in a final volume of 25μl. Primers were used to amplify the 210bp fragment of PS2 and the 304 bp fragment of cathepsin D. Amplification of 408bp of ribosomal RNA 36B4 was performed as control. The PCR mixture consisted of 1.25U GoTaq® DNA Polymerase, 1X PCR buffer (10mM Tris-HCl, 50mM KCl), 2.5mM MgCl2, 0.2mM each dNTP, 0.6μM of each PS2 primer or cathepsin D primer and 0.2μM of each 36B4 primer. PCR was performed for 20 cycles at 95°C for 1 minute, 59°C for 2 minutes and 72°C for 1 minute. Ten microliters of the PCR products were separated on a 1.2% agarose gel. (3377)

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Virology 319, 315-323. Genomic diversity of human papillomavirus-16, 18, 31, and 35 isolates in a Mexican population and relationship to European, African, and Native American variants. 2004

Calleja-Macias, I.E., Kalantari, M., Huh, J., Ortiz-Lopez, R., Rojas-Martinez, A., Gonzalez-Guerrero, J.F., Williamson, A.L., Hagmar, B., Wiley, D.J., Villarreal, L., Bernard, H.U., and Barrera-Saldana, H.A.

Notes: GoTaq® DNA Polymerase was used to amplify a portion of Human Papillomavirus 16 (HPV 16) from cervical carcinoma-derived genomic DNA. The amplified region of HPV 16 was then purified and used in sequencing reactions. (3358)

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Gene Ther. 11, 1399-1409. Highly specific transgene expression mediated by a complex adenovirus vector incorporating a prostate-specific amplification feedback loop. 2004

Woraratanadharm, J., Rubinchik, S., Yu, H., Fan, F., Morrow, S.M., and Dong, Y.J.

Notes: GoTaq® DNA Polymerase was used to amplify Green Fluorescent Protein and β-actin cDNA. This paper describes use of a “Touchdown” PCR procedure for amplification of the β-actin cDNA. Gels are presented displaying the amplification products from the reporter constructs and cells used in the experiments. (3129)

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Oncogene 23(45), 7517-7526. Nuclear insulin receptor substrate 1 interacts with estrogen receptor alpha at ERE promoters. 2004

Morelli, C., Garofalo, C., Sisci, D., del Rincon, S., Cascio, S., Tu, X., Vecchione, A., Sauter, E.R., Miller, W.H. Jr. and Surmacz, E.

Notes: COS-7 cells were harvested and total RNA isolated 24 hours after transfection with plasmids expressing trans-activating proteins. Five micrograms of RNA was reverse transcribed with oligo(dT) primer using the M-MLV Reverse transcriptase. One tenth of the reaction volume (2μl) was then used as template in PCR to amplify a gene presumed to be controlled by the trans-activators (pS2; 210bp) or a control gene (36B4 ribosomal phosphoprotein; 408bp). The PCR products were amplified using GoTaq® DNA polymerase. (3203)

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