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A New C-Terminal GST Vector for Protein Production in S. pombe
... a multiple cloning site (MCS), a GST protein-tag sequence and the nmt1 transcription termination sig...nient removal of the GST tag after purification, a sequence coding for therecognition and cleavage sites for enterokinase and thrombin have beenengineered betwe...Purified Single-Stranded M13 DNA for Automated Sequencing
... both templates. There were three and four sequence ambiguities scored for the StrataPrep- and SGB-purifed templates, respectively, within the first 20 nucleotides. Sequence ambiguities also occurred at nucleotides 553 and 638 for the StrataPrep-purified template and at nucleot...New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
...ions, and intermediatemetabolism. 1 As efforts to sequence whole genomes result in data forvast numbers of ne...ntains a DNA encoding nuclear localization sequence from the SV40 large T-antigen (amino acid PKKKRKV), amino acids 364 to 550 of the mouse NF- ...Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
...library clones ranged within ACFs expected sequence size. We successfully isolated ACF cDNA, w... spectrometric techniques provide valuable peptide sequence information from very limited amounts of protein, which can be used to screen the we...A Versatile Power Supply for All Electrophoresis Applications
...Away system and precast gels to obtain sequence from M13mp18 DNA template using the M13 (-40) primer. Sequencing results obtained using the SmartPower 4000 power supply and the CastAway system exhibit crisp, clear bands. Read lengths of approximately 400 bases per gel with ......The regions indicated as arms are the stretches of sequence homology withpAdEasy-1 vector where homologous rec...Shuttle-CMV-GFP vector was prepared by cloning the sequence for greenfluorescent protein (GFP) between the Bgl II and Xho I sites intopShuttle-CMV vector. The p...Detect Single Nucleotide Sequence Changes with Mx4000 Molecular Beacon Allelic Discrimination Kits
... result, threepossible allelic combinations of two sequence variants (Allele 1/Allele 1,Allele 2/Allele 2, All...ost-PCR manipulation ofsamples. Single nucleotide sequence changes (substitution, deletion, or insertion) arethe largest source of human DNA diversity, with an...An Epitope Tagging Vector for Gene Expression in Mammalian Cells
...ging of the protein of interest. A Kozak consensus sequence of GCC(A or G)CCATGG 7 provides optimal expression of the fusion protein when the N-terminal FLAG epitope tag is used. Other cloning options, which require fusion proteins to include their own translational start sequence, ...Automated Fluorescent-Tag Cycle Sequencing
... instrument can automatically determine thecorrect sequence of the reaction template with greater than 98% acc...intensities, whichare difficult to scan. Thus, the sequence of the resulting cycle sequencingproduct cannot be determined accurately. figure 1 ...Control RT-PCR Primers for Human Gene Transcripts with Varying Abundance
...w approach generatedlarge collections of expressed sequence tags (ESTs) that did not correspond toknown genes. The impact of this technology on new gene discovery has beenenormous. Similarly, the generation of normalized and subtracted cDNA libraries hasprovided material for the discovery of new genes. Alter...High-Level Expression of Peanut Allergens Affected by Rare Codon Usage
...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 tRNAs within a messageencoding a protein made up of 157 amino acids. This might explain theinefficient translation (...Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
...lic steroids, the EcR protein and EcRE recognition sequence weremodified to create both a synthetic ecdysone-i...the reporter pEGSH-luc in which the coding sequence for the firefly luciferase gene is inserted into the MCS of pEGSH. In these experiments, rep...Genotyping Single Nucleotide Polymorphisms with Molecular Beacons
... SNPs. SNPs are the most frequently found DNA sequence variations in the humangenome (about 1 per 350 bp ..., while the intervening loop is complementary to a sequence withinthe amplified product. When the unhybridized probe is in solution, it adopts ahairpin structur...Epitope-Tagging Vectors for Functional Analysis in Yeast
...loning site (MCS); and a transcription termination sequence downstream of each promoter. LEU2, TRP1, ...pecifically recognized by monoclonal antibodies. A sequence for the FLAG epitope (DYKDDDDK 7 ) is located in the MCS downstream of...Safe Production of High-Titer Retrovirus
... separated onto three plasmids, with minimal or no sequence overlap betweenthe plasmids, the pVPack system is ...he 3 LTR. Between the 5 LTR and the coding sequence for the gene of interest resides an extended version of the viral packaging signal (......e stop codons follow the final epitope tag sequence to ensure termination of the inserted gene transcr... separated onto three plasmids, with minimal or no sequence overlap between the plasmids, resulting in the generation of replication-incompetent ...New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
...tein purification and native eukaryotic amino acid sequence Quinn Lu John Bauer Stratagene Cloning Systems, Inc. The ESP LIC cloning and expression system features rapid and high-yieldproduction of recombinant proteins in yeast. This system usesligation-independent cloning (LIC) for efficient and direct...Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
...ated from human blood, we amplified simple sequence repeat markers. This novel isolation procedure ...igned to amplify targets containing simple sequence repeats on chromosomes 1 through 22 plus X and Y were used for this analysis (Human MapPairs...Optimizing Transfection Conditions for Studying Signal Transduction Pathways
... reporter plasmid, whichcontains the entire coding sequence of the luciferase gene downstream from abasic promoter element (TATA box) joined to five tandem repeats of the 17-bpGAL4 binding element, was used in the experiment to optimizing the trans-reportingsystem. The cis-reporter plasmid was used in experim...Production of Full-Length, Biologically Active MEK1WT and MEK1CA
... 10 For expression in E. coli, the entire coding sequence of MEK1 CA was inserted at the BamH I site of the pGEX-2T plasmid. Epicurian Coli XL1-Blue cells containing pGEX-MEK1 CA were induced by IPTG (1 mM) for4 hours, when OD 600 reached 0.6. Cells were then collected and brokenby sonication in PBS cont...High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
...orrect base insertion, as evidenced by DNA sequence analysis, and restoration of gag-pol gene function was verified by assaying reverse transcriptase activity and production of infectious virus (data not shown). Table 1 Successful Mutagenesis of a Large Plasmid with the QuikCha...A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
...Sequetech Corporation. High-signal, low-background sequence data was obtained with all four plasmid pr...parations. Figure 4 shows representative sequence data for StrataPrep endotoxin-free midiprep purified DNA. Sequence read length for the four ...High-Throughput Isolation of Total RNA
...The beacon hybridizes quantitatively to the target sequence and emits fluorescence in this conformatio...ersely proportional to the concentration of target sequence in the reaction. 6 This low-abundance target was readily detected from an RNA sampl...Organize Your Data from the Big Blue Mutation Assay
..., mutant information is entered. After DNA sequence information is available, the specific mutation(s) identified for each mutant can be cataloged and categorized in detail ( Figure 2 ). Figure 2 Reporting When data entry is complete, a range of repor...A New PCR-based Mycoplasma Detection Method
... Inasmuch as these targets share a large degree of sequence homology, bothwithin the mycoplasma genus and wi...n rate was sufficientlyrapid to contain regions of sequence divergence that could be used to speciatemycoplasmas. Once the target gene was identified, we examin...High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
...ion vectors that allow fusion of theprotein coding sequence of interest to short peptide sequences, or largerp...te, and thuspredetermine the N-terminal amino acid sequence of the protein of interest. We recently described the CBP fusion vector pCAL-n-EK 4 in whichthe li...Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
...cer ( figure 3 ). In this data set, only 3 sequence ambiguities were identified out of approxi... Only 8 ambiguities were seen in 795 bases of sequence from plasmid that was purified using the StrataPrep plasmid miniprep kit (data available ...CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
...nalysis System to obtain more than 16,000 bases of sequence dataor to analyze 1,440 genotypes per gel. DNA sequence analysis remains an essential technology for elucidating genestructure and function. For more than a...A New Lambda Vector for Mammalian Expression
... its parental pCMV-Script vector by a 29-bp sequence downstream of the f1 origin; however, this additional sequence is in a nonfunctional region of the vector. Thus, the new pCMV-Script-EX vector is functiona...Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
...h an exactsequence match to each of the two target sequence variants, genotype DNAcontrols (wild type/mutant h... out the amplification in the ABI Prism 7700 sequence detector. The genotype of a DNA sample is determined by measuring the endpoint fluorescence ...Versatile Reporter Vectors for Monitoring Viral Transduction
...approximately 8.0 kb because there is noextraneous sequence between the LTRs. We excluded selectable markers o...ia GFP (EGFP) (data not shown). The coding sequence for hrGFP was inserted between the EcoR I and Xho I sites in the pFB vector to produce the p...Yeast Protein Production System Features High Yields and One-Step Purification
...am GST gene. Successful cloning can beconfirmed by sequence analysis. To establish the expression strain, the verifiedclone is then transformed into the competent cells of S. pombe host strainSP-Q01 ((leu1-32h ) that are included in the kit. These competent cells provideoptimal transformation conditions. Tran...Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
.... Thus, recombinantproteins with native amino acid sequence are produced. Stratagene's purifiedEnterokinase is...terokinase figure 2 Coding sequence for the protein c-Jun N-terminal kinase (JNK) 3 was PCR amplified using a 5'-oligonucleotid...Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
...umn purification of protein with native amino acid sequence Denise L. Wyborski * John C. Bauer * Barbara M...amless cloning junctionsbetween the protein coding sequence of interest and the recognition target forthe site-specific protease, enterokinase (EK). The EK targ...Four New CastAway Precast Gels
...lyacrylamide gels and allow you toeither read more sequence or complete short extensions faster. The speed andconvenience of the CastAway system, combined with the longer read capabilitiesof the Long Ranger solution, make the Long Ranger gels for the CastAway systemideal for high-resolution, peak-performance ...Fractionating DNA Fragments Generated by Differential Display PCR
...ger DNA fragments are morelikely to contain coding sequence for the differentially expressed gene ofinterest, whereas DNA fragments of less than 500 bp may contain only 3 -untranslatedsequences, making identification of the gene difficult. DDPCR: Allergic and Nonallergic Blood Donors figure 2 ...The GeneZoo or the GeneJungle?
...ame, keyword, UniGene ID, or nucleotide or protein sequence (www.stratagene.com). A gel image of a restriction...an untapped resource for new discoveries. The raw sequence of the human genome is now mostly complete, but it will be quite some time before all of the genes a...Constructing Directional cDNA Libraries from Limited Amounts of RNA
...hough a shift occurred in the average insert size, sequence analysisrevealed that the distribution of gene cla...hesized, then ligated to an adaptorthat contains a sequence for LIC. The adaptor-ligated cDNA is amplified by PCRusing primers with specifically designed LIC en...Randomize Gene Sequences with New PCR Mutagenesis Kit
...s, including a 2.5-kb polymerase gene and a 720-bp sequence encoding Green Fluorescent Protein (GFP). ...ymerase. These data were compiled from DNA sequence analysis (37.5-kb total) of 45 clones randomly selected from lacZ mutagenesis studies. Mutat...Generate High-Quality, Directional Plasmid cDNA Libraries
...raditionally, genes have been selected for sequence homology; more recently, genes are being d...ing, complementation studies and expressed sequence tag (EST) sequencing, cDNA libraries constructed directly into plasmid vectors are convenien...High-Fidelity PCR with a Novel Polymerase Mixture
...luding forward mutation assays, 4,5,6 DNA sequence analyses 7 and denaturant gradient gel electrophoresis (DGGE) analyses 8 have been used to show that Pfu DNA polymerase has the lowest error rate of any thermostable DNA polymerase studied to date. Pfu DNA polymerase also la...High-Titer Retroviral Vectors for Gene Delivery
...he vector pFBhas been deleted for all viral coding sequence and contains no exogenoussequence other than that ...are based on MMLVand contain an extended packaging sequence and splice-site configuration thatincreases viral titer and gene expression from the viral promoter....HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
... structureand contain approximately 50% nucleotide sequence identity. Current diagnostic methods for HSV infe...cture. 4-7 The loop is comprised of a DNA sequence complimentary to the target DNA, and the stem is formed by short complimentary sequences at ...Improved Vectors for PathDetect Trans-Reporting Systems
...fthe activation domain of any transcription factor sequence and subsequentexpression of GAL4 fusion proteins i... that does not contain the GAL4 DNA binding domain sequence and doesnot express protein from the CMV promoter. The pFR-Luc plasmid (0.5 g) and pFC-MEKK...High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
...R kit resulted in highly purified DNA with sequence data showing low-background, high-signal intensity...rast, the QIAquick 96 PCR kit demonstrated sequence data that had high background, variable signal intensity, and numerous ambiguities. ...Mammalian Expression Vector for Efficient Cloning of PCR Fragments
...T4 DNA ligase. The Srf I enzyme recognizes therare sequence 5 -GCCCGGGC-3 .By including Srf I enzyme in the ligation reaction, the PCR-Scriptcloning method maintains high concentrations of digested vector DNA and allowsthe use of nonphosphorylated, unmodified PCR products. The ligation efficiencyof blunt-en...Easy Cosmid DNA Purification with StrataPrep Kits
... DNA. A representation of the high-quality sequence is shown in Figure 3 . The read length was approximately 630 bases with only four sequence ambiguities. Fig.3 The StrataPrep plasmid minipr...New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways
...midcontains direct repeats of the gamma-activating sequence from the promoter ofthe guanylate-binding protein....omodimer, which binds to theunique gamma-activated sequence (GAS), is formed in response to IFN- g . 3 The mechanism of the activation and nuclear localization ...Sequence-Validated and Expression-Tested Human cDNA in a Dual Expression Vector
...om. Validated Nucleotide Sequence The nucleotide sequence of the ORF of all expression-tested clones has bee...ing. A comparison between the clone and apublished sequence is provided with each clone. Epitope and Purification Tags All clones express a fusion prote...New Fusion Trans-Activator Plasmids for Studying Signal Transduction Pathways
...ivation domain of any transcription factor sequence and subsequent expression of GAL4 fusion proteins in a variety of mammalian cells. The pFA-CMV plasmid features (1) a multiple cloning site (MCS) that offers 10 unique, conveniently arranged restriction sites for insertion of ...Stratagenes Human cDNA Microarrays: Perform Gene Expression Profiling and Discover New Genes
...from the 3 end ( Figure 2 ). The 3 sequence information is then used to design an insert-specific primer that is capable of amplifying approximately 350 bases of the 3 end of the cDNA, including the polyA tail. After PCR with the insert-specific primer, the pres...Fast Isolation of Total RNA from Small Numbers of Cells
...ted, no bands were detected; therefore, the target sequence DNA did not contaminate the reactions. ...ion. The molecular beacon hybridizes to the target sequence and emits fluorescence in this conformation. The threshold cycle number (C t ) is in...Cis-Reporting System Monitors Intracellular Calcium Mobilization
... have distinctaffinities, they bind to a consensus sequence of T(A)GGAAAAT(A/C). 1,2 This site is present in m...th four direct repeats of the NFAT binding sequence (286 to 257) from the IL-2 gene promoter, the most studied and widely used NFAT binding ...Cis-Reporting System Monitors Intracellular Calcium Mobilization
... have distinctaffinities, they bind to a consensus sequence of T(A)GGAAAAT(A/C). 1,2 This site is present in m...th four direct repeats of the NFAT binding sequence (286 to 257) from the IL-2 gene promoter, the most studied and widely used NFAT binding ...EppendorfPerfect gDNA Blood Mini Kit
...ted product of amplification of the target sequence with these primers was 687 bp in length. Two other... expected amplification product of the target sequence with these primers was 168 bp in length. The PCR reaction was performed on 2.5 l of e...EppendorfPerfect gDNA Blood Mini Kit
...ted product of amplification of the target sequence with these primers was 687 bp in length. Two other... expected amplification product of the target sequence with these primers was 168 bp in length. The PCR reaction was performed on 2.5 l of e...Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
...alysis of gene expression in eukaryotes, expressed sequence tag (EST) data represent an invaluable resource for gene identification. EST's are single pass partial cDNA sequences. In general, cDNA clones are selected to represent as many unique transcripts as possible. One such ...Efficient and Reliable PCR Setup Using Eppendorf MasterMix
...xon 26), a 547-bp fragment with the coding sequence of the pyrophosphatase gene of Thermo thermophilus...) Lane 4: 547-bp fragment from the coding sequence of the pyrophosphatase gene Thermus thermophilus (70% GC), (genomic DNA) Lane 5: 10......rs to confirm the expression of the target sequence via RT-PCR in the cells under examination. If the RT-PCR is positive, the amplicon can be used as probe for detecting the corresponding mRNA on Northern blots or for semi-quantitative RT-PCR. As many sequences are low-copy gen...... of sub-cellular location), the missing 5' sequence has to be obtained at a later point in tim...rnative is direct PCR amplification of the missing sequence using the cDNA library as template [1,2]. In this case, a vector primer (VP) and a g...Other ContentsdifficultydifficultydifficultydifficultydifficultydifficultydifficultydifficultyswallowingswallowingswallowingswallowingswellingswellingswellingswellingswellingswellingswampsymptomaticsymptomaticsymptomaticsymptomaticsymptomaticswansweatingsweatinggranulomaOther TagsPolishPolishActivistsForcepsEarthquakeAirwayAirway
(Date:9/4/2008)...ful supercomputers capable of analyzing decades of...milestone capable of bringing comprehensive change...worldwide. Researchers at the University of Miami,...nce, collaborating with NCAR (National Center for ...-Atmospheric Studies) and the University of Califo...
(Date:9/4/2008)...08) A variety of organismsfrom bacteria and fung...ere the timing and duration of fundamental biologi...to adapt and survive, even under fluctuating envir... advances have been made to understand the molecul...nto modifications in cellular physiology and organ...
(Date:9/4/2008).... , Cystic kidney disease, renal fibrosis, or re... organs are characterized by overproduction or on...in the renal cells. An international research team...Heidelberg University Hospitals has now developed ...ions. , The researchers introduced a genetic sw...
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Breaking Biology News(10 mins):Petascale climate modeling heats up at University of Miami 2Petascale climate modeling heats up at University of Miami 3Interdisciplinary volume on biological rhythms serves as both primer and in-depth resource 2Gaining a better understanding of kidney diseases 2Underwater microscope helps prevent shellfish poisoning along Gulf Coast of Texas 2863 1Underwater microscope helps prevent shellfish poisoning along Gulf Coast of Texas 2863 2Underwater microscope helps prevent shellfish poisoning along Gulf Coast of Texas 2863 3For Virginia Tech Community Anxiety and Sadness May Increase on Anniversary of School Shootings 16605 1For Virginia Tech Community Anxiety and Sadness May Increase on Anniversary of School Shootings 16605 2La Jolla Institute for Allergy 26 Immunology Wows With New Website 4551 1La Jolla Institute for Allergy 26 Immunology Wows With New Website 4551 2With annual deaths from malaria on the rise 3A Scientists ask where is all the money going 3F 16602 1With annual deaths from malaria on the rise 3A Scientists ask where is all the money going 3F 16602 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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