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New Microarray Labeling Kit for Preparing cDNA Probes
...y andreproducibility of identifying differentially expressed genes. The kit includesseveral control mRNA and tw...x additional human target DNA. Thesetarget DNA are expressed in different tissues at varying abundances and provideadditional information regarding the quality o...A New C-Terminal GST Vector for Protein Production in S. pombe
...periments, the firefly luciferase gene was expressed using the pESP-3 vector. The luciferase-GST fusion protein was purified to near homogeneity with a yield of 12.5 mg/liter (data not shown). Stratagenes FLAG Western Detection Kit can be used to monitor expression an...New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
... cell. The protein of interest, or bait, is expressed as a fusion protein with the GAL4 DNA binding domain, and the target protein is expressed with the activation domain. If the bait and target proteins interact, the activation...Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
... base aided in isolating a protein that was widely expressed but only in small amounts, and the average size of the library clones ranged within ACFs expected sequence size. We successfully isolated ACF cDNA, which was crucial in allowing us to define the minimal protein require...New Kit Generates Exceptionally Pure Total RNA for RT-PCR
...e expression analysis because it amplifies expressed RNA sequences. 1 RT-PCR is very sensitive... 3 ). Similarly, ornithine decarboxylase is expressed at overall low levels but the levels vary in tissues throughout the body. 8 The pre...Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
... (RLU) were normalized toprotein concentration and expressed as RLU/mg of protein. The transfection efficiency of StrataPrep-purified DNA, measured as total RLU/mg of protein for stimulated cells, was 1.3-, 3.4-, and 2340-fold higher than stimulated cells transfected with CsCl, AEC, and ...An Epitope Tagging Vector for Gene Expression in Mammalian Cells
...malian Cells Use tag-specific antibodies to study expressed genes Tanya Hosfield Quinn Lu Stratagene The pCMV-Tag1 vector is an epitope tagging vector designed for geneexpression in mammalian cells. A target gene inserted into the pCMV-Tag1 vectorcan be tagged with the FLAG epitope (N-terminal, C-t...Brilliant Core Reagent Kits for QPCR and QRT-PCR
...pression levels. Mutations can also be detected in expressed RNA with these kits ( Table 2 ). Table 2 Comparison of QRT-PCR Kits Kit Name Enzyme dUTP/UNG Passive Reference Dye Number of rea...Control RT-PCR Primers for Human Gene Transcripts with Varying Abundance
...1 This new approach generatedlarge collections of expressed sequence tags (ESTs) that did not correspond tokno...ression analysis, housekeeping genes are typically expressed at higherlevels than the gene of interest, which makes comparisons and quantificationdifficult. To ...High-Level Expression of Peanut Allergens Affected by Rare Codon Usage
...onstrating that Ara h 2 is only marginally expressed in these conventional E. coli cells. Ara h 2 Codons Examination of the codons used in this sequence indicated that Ara h 2contains 12% of the least used codon AGG/AGA in E. coli and,additionally, 2% of other rare codons read by minor ...Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
... vector is engineered such that both receptors are expressed from a single mRNA transcribed from the CM...tains a neomycin-resistance gene, which is expressed in both E. coli (kanamycin-resistance) and mammalian cells (G418-resistance). ...Detecting Gastrin-Releasing Peptide Receptor by in situ PCR on Archived Tissue
...rointestinal (GI) tract,GRP receptors are normally expressed by smooth muscle cells and by nerveendings, acting...udies have shown that GRP receptors are aberrantly expressed by anumber of human colon cancers 6 and colon cancer cell lines includingHT29, MC26 and NCIH716. 7,...Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
...ion of FLAG -Tagged Fusion Proteins Detection of expressed tags made easy Simple, Sensitive, and Rapid Dete...s to detect a GST-FLAG-MEK1 fusion protein expressed under the control of the inducible promoter. 2 When samples derived from cells were grown u...Epitope-Tagging Vectors for Functional Analysis in Yeast
... shown in Figure 3 , all eight constructs expressed active luciferase. When pESC-TRP-LucFN or pESC-TRP-LucMC was used, under induced conditions, the activity of the epitope-tagged luciferase showed a level similar to that of the untagged luciferase ( Figure 2 ). The dat...Safe Production of High-Titer Retrovirus
...n protein and each of the envelope proteins are expressed at sufficiently high levels to allow the production of viral supernatants with titers greater than or equal to 10 7 cfu/ml following transient transfection. Safety: Because there is minimal sequence overlap between the packaging vectors ......rest. Inserts into the phrGFP-C vector are expressed as C-terminal fusions to hrGFP. We tested the protein fusion vectors by fusing hrGFP to a series of genes that encode subcellularly localized proteins. In Figure 3, each fluorescent microscope photograph shows hrGFP expressio...New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
...ication Figure 3A & 3B We have expressed and purified a number of recombinant proteins in S. pombe, using the pESP-2 vector and the LIC cloning strategy to create the constructs. The recombinant proteins include human Elk1, Krs2, MEK1 WT and the chicken calmodulin g...Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
...d for amplifying DNAtargets that are to be cloned, expressed and/or sequenced. Moreover, PfuTurboDNA polymerase generates significantly higher yields of PCR product than can beobtained with other single PCR enzymes, and the enhanced performance of PfuTurboDNA polymerase allows PCR amplifications to be conducte...Optimizing Transfection Conditions for Studying Signal Transduction Pathways
..., panel A). The GAL4-ATF2 fusion protein, which is expressed from the pFA-ATF2 plasmid, is strongly activated by the MEKK protein in transient transfection assays. 9,10 In this experiment, HeLa cells (1.5 x 10 5 /well) were cotransfected with the indicated amounts of the pFA-AT...Human Universal cDNA Library Array I
...niversal cDNA library that represent genes expressed in 29 different human tissues. This normalized library demonstrates a good average length of inserts, low representation of the ubiquitous b-actin gene, and high percentage of full-length clones. The bacterial colonies origina...Comparing Fidelity and Performance of Proofreading PCR Enzymes
...Pfu DNA polymerase. 4 DNA polymerase fidelity is expressed as error ratethe mutation frequencyper base pair per duplication (MF/bp/d). When determining polymerase error ratesin PCR-based assays, mutation frequencies [# mutants/total # clones] must benormalized with respect to the number of template doublings...Production of Full-Length, Biologically Active MEK1WT and MEK1CA
...loning Systems, Inc. Stratagene has successfully expressed and purified recombinant, wild-typehuman MEK1 WT ...search and drug screening. Although MEK1 WT canbe expressed in soluble form in E. coli as glutathione-S-transferase(GST) fusions, these products are largely deg...Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
...r the IL-4 receptor (IL-4 R), which is known to be expressed on the surface of Daudi cells. NIH3T3 cells were infected at an MOI of approximately one, and, after 2 days, the cells were labeled with receptor-specific monoclonal antibody followed by a fluorescent anti-mouse IgG secondary ...RT-PCR Primer Sets for Human and Mouse Mismatch Repair Genes
...enetic integrity of thecell, they are likely to be expressed in detectable levels in most cell types.In this regard, they could be classified as housekeeping genes. Thus, the RT-PCRprimer sets could be used as positive controls for cDNA templates in RT-PCRstudies unrelated to mismatch repair. PCR Amplificatio...High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
... high-level production and one-steppurification of expressed proteins. Similar to its predecessor pCAL-n-EK,pCA...d in E. coli, and CBP fusionproteins are typically expressed to high levels. In addition, the CBP tag isefficiently phosphorylated by protein kinase A and thus, ...Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
...xperiments identified several differentially expressed genes in the invasive rim of the tumor relative to the noninvasive tumor core. 4 Conclusions The StrataPrep total RNA microprep kit is ideal for the isolation of RNA from small cell samples harvested by LCM. The method ...A New Lambda Vector for Mammalian Expression
...mid vector for analysis of transcripts via expressed sequence tag (EST) sequencing or subtraction and normalization procedures. The HybriZAP 2.1 vector offers the convenience of lambda library technology; after library construction, excision of the pAD-GAL4-2.1 plasmid vector al...Yeast Protein Production System Features High Yields and One-Step Purification
...that can be critical for thebiological activity of expressed proteins. By using the yeastSchizosaccharomyces po...protein in the pESP-1 vector that has been expressed in cells grown under repressed and induced conditions. The fusion protein cannot be detected...Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
...catalytic subunit of bovine EK has been cloned and expressed inE. coli and purified to homogeneity. 1, 2 Strat...on tag. The resulting fusion protein, CBP-JNK, was expressed in Epicurian Coli BL21(DE3) cells and purified from crude cell lysates to apparent h...Using XL1-Red Mutator Strain to Generate Esterase Variants
...oding a PFE from Pseudomonasfluorescens, which was expressed well in E. coli using arhamnose-inducible promoter. Since the structure of PFE is not known, positionsfor site-directed mutagenesis are difficult to predict. Alternatively, a randommutagenesis of the gene combined with an assay system also allows det...Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
... high-level productionand one-step purification of expressed proteins. A ligation-independent cloning(LIC) stra...de (CBP) purification tag and the N terminusof the expressed protein. Because EK cleaves at the C terminus of itsrecognition target, which is also the N terminus...pFB-ERV: Retroviral Delivery of the Ecdysone Receptor Proteins
... transfection assays. The neo-resistance marker is expressed as thethird open reading frame (ORF) in a tricistr... 1 ). The receptor proteins VgEcR and RXR are expressed from the first and second open reading frames (ORF), respectively, and the neomycin-resista...Fractionating DNA Fragments Generated by Differential Display PCR
...used technique foridentifying genes differentially expressed between cell types. Application ofDDPCR to allergi... methods may be used for identifyingdifferentially expressed genes, such as high-throughput sequencing of expressedsequence tags (ESTs), subtractive hybridizatio...Constructing Directional cDNA Libraries from Limited Amounts of RNA
...with the expected sequences that we observed to be expressed in liver tissue. Table 1: Breakdown of Sequencing Results by Class Class 1: Sequence is identical to a known human gene Class 2: Sequence has a significant match with a human protein Class 3: Sequence has a signific...Generate High-Quality, Directional Plasmid cDNA Libraries
...rid screening, complementation studies and expressed sequence tag (EST) sequencing, cDNA libraries constructed directly into plasmid vectors are convenient and easy to use. However, constructing high-quality plasmid libraries can be limited by the efficiency of the cDNA synthesi...Gene-Specific Primers for NMDA Receptor Subunits
... protein can form NMDA-activated ion channels when expressed in Xenopusoocytes, but the currents in such channels are much weaker than those seen insitu. Channels with more relevant physiological characteristics are producedwhen the NR1 subunit is combined with one or more of the NMDAR2 (NR2A-D)subunits. 3 In...Codon Bias-Adjusted BL21 Derivatives for Protein Expression
... for high-level production ofheterologous proteins expressed from the T7-promoter. Thesestrains have been eng...recombinant genes that are only marginally expressed in conventional BL21 strains, BL21-CodonPlus(DE3)-RIL cells were transformed with vectors ex...New BL21-CodonPlus Cells Correct Codon Bias in GC-Rich Genomes
...sed as hosts for expressing genes that are expressed well in conventional E. coli strains. Expression of Genes from GC-Rich Organisms To confirm the function of the argU and proL tRNAs in BL21-CodonPlus(DE3)-RP cells, we performed two test transformations ( Figure 2 ). BL21-Codo...hNT BlueNeurons Express b-Galactosidase at High Levels
...sults indicate that the beta-galactosidase gene is expressed at a sufficient level to yield intense blue staining of the transfected neurons. Conclusions hNT BlueNeurons have high levels of beta-galactosidase activity and stain dark blue in standard assays. This feature is especially useful f...High-Titer Retroviral Vectors for Gene Delivery
...lot (1:10 5 for both viruses). Titers are expressed as the product of the dilution factor multiplied by the percentage of infected cells that fluoresce above background, then multiplied by the number of cells infected. By this definition, pFB-Neo-GFP infected cells showed a tit...Identification of Differentially Expressed Gene Products with the CastAway System*
...gels were used to detectand isolate differentially expressed gene products amplified by the RNAarbitrarily prim....Fourteen unique bands representing differentially expressed gene products due tothe PMA stimulation were identified, isolated from the gel and reamplified byPCR...TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins
...er University,Ontario, Canada GST-fusion proteins expressed in TKB1 bacteria are used to affinity purifycellul...1 strain, yet efficiently bound gT-YD when expressed in TKB1. The lack of binding can neither be attributed to the amounts nor the level of tyros...Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
...cer. 1 The MSH2 gene encodes a 3.1-kb mRNA that is expressed at low to moderate levels. 2 The larger-than-average size and modest expression level of this RNA make it agood candidate for evaluating products for Northern blot analysis. Fig.1 Stratagenes products for o...RT-PCR Primers for the Study of Apoptosis
...er, the Fas antigen gene is differentially expressed in certain tissues. No amplification occurred in the control rat and human cDNA libraries. Figure 4 Rat Fas Antigen The rat Fas antigen gene was amplified in rat brain, lung, and heart cDNA libraries (...A Comparison Study of Lipid Tranfection Reagents in A549, NIH 3T3 and COS-7 Cell Lines
...totoxic effects of either the lipid or the expressed plasmid protein. The greater the amount of lipid tolerated by the cell line, the more transfection is achieved. To maximize the amount of lipid, we compared the concentration of cell proteins after transfections (data not show...Mammalian and Bacterial Expression in One Vector
...ream ofthe cloning site. The 4-kDa CBP fusion tag, expressed on the C-terminus of theprotein, can then be used for easy detection and purification of the fusionprotein using Stratagenes Affinity CBP fusion detection kit and Calmodulin Affinity Resin, respectively. The newpDual expression vector is offered e...High-efficiency Derivatives of the BL21 Series for Protein Expression
...ue dye. For the JNK protein, the amount of protein expressed from both strains of competent cells was indistinguishable. Expression of JNK is shown in Figure 2 . Figure 2 Conclusions The BL21-Gold series of competent cells allows researchers to directlytransf...Sequence-Validated and Expression-Tested Human cDNA in a Dual Expression Vector
...atile dual-expression vector permitsproteins to be expressed and detected in mammalian cells and proteins to be... for convenient and sensitive detection of expressed proteins with anti-c-myc antibody. The HIS6 purification tag consists of six histi...New Fusion Trans-Activator Plasmids for Studying Signal Transduction Pathways
...in activates the GAL4-c-Fos fusion protein expressed from the pFA-cFos plasmid. The mechanism o...ith previous reports, the GAL4-ATF2 fusion protein expressed from the pFA-ATF2 plasmid was strongly activated by the MEKK protein in transient tr...High-Efficiency Transfections Achieved with New Low-Toxicity Reagent
...ain cells with X-gal, and the resulting cells that expressed for b-galactosidase were photographed ( Figure 2 ). Fig.2 Assessing Post-transfection Cell Viability We evaluated the cytotoxic effects of GeneJammer and the competitive reagents by counting the cells staine......f MA, Tan C, Conrad LJ and Klein PS. Frizzled-8 is expressed in the Spemann organizer and plays a role in early morphogenesis. Development. 1998;125:2687-2700. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. 1989;3:A1-A3. ibid.......nd that the gene is not already endogenously expressed in the examined cells. Reporter systems are also often used as a standard to compare the transfection efficiency of different transfection experiments. In this case, the control reporter system usually contains a constitutive p......nd that the gene is not already endogenously expressed in the examined cells. Reporter systems are also often used as a standard to compare the transfection efficiency of different transfection experiments. In this case, the control reporter system usually contains a constitutive p...... functions (phase I and phase II enzymes) are expressed for up to 10 days in culture under optimal conditions. Fig. 4: Optimal signal to background ratio Hybridization of a Rat Liver Chip on CreativeChip 3D Slide with Cy3-labeled cDNA from 100 g...Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
...For the analysis of gene expression in eukaryotes, expressed sequence tag (EST) data represent an inval... (Cye5) spots indicate that the targets are expressed differentially. Figure 3 The 1,152 cDNA clones derived from the mouse Un...Enhancing PCR* Efficiency with MasterTaq
...er confirmation that they really have been expressed differentially. One way of achieving this aim ...s MB. Identification and cloning of differentially expressed genes. Curr Opin Biotechnol. 1997;8(5):542-6. Wang Z and Brown DD. A gene express...Normalizing cDNA libraries using the EppendorfThermomixer
...to over-representation of clones of highly expressed genes, whereas "least abundant" mRNAs can only be ...sent: 1. Rehybridized ds-cDNAs from highly expressed mRNAs 2. Ss-cDNAs from rare mRNAs 3. Partly rehybridized cDNAs from frequently-occ...Effectene Transfection Reagent provides efficient gene delivery to primary neuronal cell cultures
...sfection conditions. Transfection efficiencies are expressed as the percentage of neurons that are GFP-positive. Note that transfection efficiency decreased when more than 5 l of Effectene Reagent was added. Highest transfection efficiencies were obtained using 200 ng of DNA in combination ...Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
...neuronal and glial cells. The cultured cells expressed characteristic markers and showed typical morphology and differentiation. We transfected the cells with two plasmids, one encoding green fluorescent protein (GFP) and the other encoding -galactosidase (-gal). No obvious signs of tox...... Average transfection efficiencies are expressed as percentages relative to the efficiency obtained with DNA prepared using the QIAGEN Plasmid Kit (100%). Plasmid Purity vs. Transfection Efficiency...Other ContentsneuropathyneuropathyneuropathyneuropathysmearsmearsmearsmearperitonitispersonalitypersonalitypersonalitypersonalitypertussispertussisperitonsillarpersonalpersonalpersonalpersonalpersonalpersonalpersonalpersonalpersonalpesticidespesticidespesticidespesticidesOther TagsFebFAKFAKOther Tagspermissible 2buying 2buying 3buying 4buying 5buying 6buying 7mere 2mere 3mere 4mere 5islands 2islands 3vegetation 2vegetation 3
(Date:9/3/2008)...ush has named the recipients of the 2007 National ...cience and engineering. Honorees will receive the .... , "The 2007 National Medal of Science laureate...superb research," said Arden L. Bement, Jr., direc...ir legacies have impacted our lives in numerous wa...
(Date:9/3/2008)...ists of places with the worst air pollution in the...proved the situation, tiny particles called aeroso... from UC San Diego and six other institutions have...to precisely identify aerosols that make up the ha... , This forensic work will help to identify the ...
(Date:9/3/2008)...olecular and evolutionary biologists in collaborat...the full genome sequence of Trichoplax , one of n...oviding a new insight into the evolution of all hi...ine edition of the journal Nature show that whil...omes found in a multi-cellular creature, it contai...
(Date:9/3/2008)...ge flightless birds of the southern continents Af...South American rheas and the New Zealand kiwi do ...ieved. , Instead, each species individually lost...d have the ability to fly, according to new resear...ogy professor Edward Braun. , The new research, ...
Breaking Biology News(10 mins):White House announces 2007 National Medal of Science laureates 2Smoke smudges Mexico City's air, chemists identify sources 2Smoke smudges Mexico City's air, chemists identify sources 3Trichoplax genome sequenced -- 'rosetta stone' for understanding evolution 2New research challenges long-held assumptions of flightless bird evolution 2Study Supports Possible Use of Leukine 28R 29 as a Potential Adjuvant Therapy for High Risk Melanoma Patients 2335 1Study Supports Possible Use of Leukine 28R 29 as a Potential Adjuvant Therapy for High Risk Melanoma Patients 2335 2Study Supports Possible Use of Leukine 28R 29 as a Potential Adjuvant Therapy for High Risk Melanoma Patients 2335 3Study Supports Possible Use of Leukine 28R 29 as a Potential Adjuvant Therapy for High Risk Melanoma Patients 2335 4Study Supports Possible Use of Leukine 28R 29 as a Potential Adjuvant Therapy for High Risk Melanoma Patients 2335 5Milestone Scientific Expands U S and Canadian Distribution Network With Addition of Patterson Dental Supply 21174 1Milestone Scientific Expands U S and Canadian Distribution Network With Addition of Patterson Dental Supply 21174 2Milestone Scientific Expands U S and Canadian Distribution Network With Addition of Patterson Dental Supply 21174 3Milestone Scientific Expands U S and Canadian Distribution Network With Addition of Patterson Dental Supply 21174 4Bipolar Teens at Greater Risk of Substance Abuse 21171 1Bipolar Teens at Greater Risk of Substance Abuse 21171 22008 Gruber Genetics Prize awarded to Allan Spradling of Carnegie Institution 21167 12008 Gruber Genetics Prize awarded to Allan Spradling of Carnegie Institution 21167 2(Date:9/4/2008)... may be shortchanging their golden years by tappin...iversity of Illinois expert who has studied the fe..., More than three of every four Americans trim th...iming benefits early, a choice many might forego i... consequences, law professor Richard L. Kaplan say...
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Breaking Medicine News(10 mins):Health News:New guide explores making the most of Social Security 2Health News:New guide explores making the most of Social Security 3Health News:Oncology Clinical Research Scientist Recommends a Nuanced Preclinical Study Process to Speed Drug Development 2Health News:Oncology Clinical Research Scientist Recommends a Nuanced Preclinical Study Process to Speed Drug Development 3Health News:Schofield Publishing Enters the Newsletter Market 2[0] New guide explores making the most of Social Security[0] Oncology Clinical Research Scientist Recommends a Nuanced Preclinical Study Process to Speed Drug Development [0] Schofield Publishing Enters the Newsletter Market[0] ULURU Inc. to Present at the Thomas Weisel Partners Healthcare Conference 2008
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