A New C-Terminal GST Vector for Protein Production in S. pombe
...translation start site, a multiple cloning site (MCS), a GST protein-tag sequence and the nmt1 transcription termination signal. The nmt1 promoter of S. pombe is tightly repressed in the presence of thiamine (vitamin B 1 ) in the growth medium and is highly activated upon i...A Positive Selection Assay for Mutation Analysis in Big Blue Animals
... in length, including the cII mRNAribosome binding site and the cII protein-activated P RE promoter. 5,6 The relatively short length of the cII target region simplifiessequencing and expedites the analysis of mutational spectra. Conclusions The l Select-cII mutation assay kit offers analternative to t...Assess the In Vivo Activation of Signal Transduction Pathways with PathDetect Reporting Systems
...ver, the pFA-CMVvector features a multiple cloning site with 10 unique, conveniently arrangedrestriction s...ption factor of interest into the multiple cloning site of the pFA-CMVvector, researchers will have their own fusion trans-activator vectors forstudying sig......he pShuttle-CMV vector contains a multiple cloning site between the CMVpromoter and the SV40 polyadenylati...he pShuttle vector contains onlya multiple cloning site in which an entire expression cassette can be inserted.The regions indicated as arms are the stretch...An Epitope Tagging Vector for Gene Expression in Mammalian Cells
...esistance in mammalian cells. The multiple cloning site (MCS) of the pCMV-Tag1 vector is arranged to allow a variety of cloning strategies to be used, resulting in C-terminal, N-terminal or internal tagging of the protein of interest. A Kozak consensus sequence of GCC(A or ...Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
...nous host genes, as well as asynthetic recognition site that would not be recognized by host transcription...reporters containing the synthetic binding site AGTGCA N1 TGTTC (E/GRE). This binding site is extremely unlikely to be recognized by steroid...Genotyping Single Nucleotide Polymorphisms with Molecular Beacons
...ular beacon design is available on Stratagenes web site at http://www.stratagene.com/index.asp?catID=17. The nucleotide sequence of the molecular beacons was as follows: SNP11A: 5 -fam-gcgca gagacggAacttgctctgaaa ctgcgc-DABCYL-3 ; SNP11G: 5 -fam-gcgca gagacggGacttgctctgaaa ctgcgc-DABCYL-3 . Underlined...Epitope-Tagging Vectors for Functional Analysis in Yeast
...n opposite orientation; a multiple cloning site (MCS); and a transcription termination sequence ...ciferase gene was individuallycloned in the BamH I site or the Not I site of pESC-TRP, so thatthe gene is under the control of the GAL1 promoter or the GAL1...Safe Production of High-Titer Retrovirus
...RFs) are all followed by an internalribosome entry site (IRES) linked to a downstream drug-resistance gene... of the MMLV genomic DNAand deleting the packaging site so the gag-pol and env-expressinggenome could not be packaged itself. However, a problem inherent wi......vectors (Figure 1) have a multiple cloning site upstream of three contiguous copies of eit...by the coding region of an internal ribosome entry site (IRES) for reentry of the ribosome and protein production of the downstream hrGFP ge...New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
...nsferase (GST)purification tag and the recognition site for enterokinase. After expression andsingle-colum...ting in aseamless junction between the EK cleavage site and the initiation codon (ATG) ofthe gene to be expressed. After expression in S. pombe, the GST fus...Human Universal cDNA Library Array I
...poly(C) and at their 3 termini by an Sac I site (reconstituted by the cohesive ends of the primer/adapter and vector). These molecules were transformed into E. coli resulting in 2 x 10 5 ampicillin-resistant clones. Figure 1 The remaining cDNA...Production of Full-Length, Biologically Active MEK1WT and MEK1CA
...ing 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 containing 1% Triton X-100 and protease inhibit...Restriction Endonucleases Undergo Rigorous QC Testing to Ensure Optimal Performance
... exonucleasesthat destroy the integrity of the cut site by cleaving nucleotides from theresultant 5 or 3...minants The integrity of a restriction enzyme cut site can be destroyed by thepresence of exonucleases that can digest single-or double-stranded ends.Strat...Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
... 1A ). The pFB vector has an extended packaging site and uses the viral splice donor and splice acceptor configuration from the high-titer, high-expressing vector pMFG. 7 To minimize packaging size constraints, and because the high infection efficiency of the system obviates th...High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
... proteases that cleave internal to the recognition site and thus,leave at least one extraneous amino acid ...kinase cleavage site, the multiple cloning site (MCS), and the T7 transcriptional terminator. LIC Cloning of JNK ...Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
...s, each containing a different restriction site sequence. The PCR product was digested with both restriction enzymes, purified using each method, ligated to digested pBluescript SK(+) vector and transformed into XL2-Blue ultracompetent cells. In five separate experiments,...A New Lambda Vector for Mammalian Expression
... the firefly luciferase gene into the BamH I site of the MCS. This same luciferase gene was inserted into the parental pCMV-Script vector. These two constructs were used for parallel transfections of Chinese hamster ovary (CHO) cells. Cell lysates were prepared and assayed fo...Yeast Protein Production System Features High Yields and One-Step Purification
...T protein-tag sequence, a multiple cloning site and the nmt1 transcription termination sig... of the gene of interest into the multiple cloning site results in an N-terminal fusion with the GST peptide, which facilitates one-step pur...Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
...esigned to fuse the 5-amino-acid EK target site to the natural amino terminus of the JNK protein. The PCR product was inserted into the affinity pCALnEK expression vector 4 such that the EK target peptide-JNK fusion protein was fused in frame, downstream of the calmodulin-...Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
... a seamless junction 4,5,6 between the EKcleavage site and the protein coding sequence. The protein sequence isefficiently cloned into the pCAL-n-EK vector, and protein expression is inducedby the addition of isopropyl-thio-D-galactoside (IPTG). The fusion protein ispurified in one step using the CBP aff...Constructing Directional cDNA Libraries from Limited Amounts of RNA
...esistance in mammalian cells. The multiple cloning site (MCS) of the pCMV-PCR vector is flanked by T3 and T7 promoters, which allow generation of RNA by in vitro transcription of a DNA insert in either orientation. Fig.1 Ovarian Tumor Library An ovarian tumor library...Generate High-Quality, Directional Plasmid cDNA Libraries
...-primer that contains an Xho I restriction site near its 5 end. First-strand synthesis t...om digestion, only the nonmethylated Xho I site in the linker-primer is cleaved. The resulting cDNA contains an Xho I-compatible overhang ...High-Titer Retroviral Vectors for Gene Delivery
...ssequence other than that for the multiple cloning site (MCS). As has been shownfor other such minimal MML...ved by extending the 3 border of the Y site further into the gag coding sequence relative to other MMLV vectors. 5 This extended ...HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
... can bereactivated to cause lesions at or near the site of initial infection. 1 Most common are mucocutaneous infections, which primarily affect oral-facial andgenital surfaces. However, more serious HSV infections can also involve thecentral nervous system and viscera, particularly in newborns andimmunoc...TKB1 Cells Identify Receptor Tyrosine Kinase Interacting Proteins
...which interact with a specific autophosphorylation site onthe ErbB2/Neu receptor tyrosine kinase (RTK). Th...onstrated bythe binding of SHC protein to the same site as observed in vivo. We failed todetect proteins that do not interact with these phosphorylation sit...In Vivo Signal Transduction Pathway Reporting Systems
...with a mutation in the PKA phosphorylation site of the pFA-CREB plasmid showed that the mutated fusion activator could not be activated by the PKA protein. Activation of PathDetect Reporting Systems by Extracellular Stimuli Figure 4 In addition to activation by...Improved Vectors for PathDetect Trans-Reporting Systems
...lls. The pFA-CMVvector features a multiple cloning site with 10 unique, conveniently arrangedrestriction sites for insertion of any activation domain sequence; the CMVpromoter for high-level expression of fusion trans-activator proteins inmany mammalian cell lines and the neomycin-resistance gene, which f...Mammalian and Bacterial Expression in One Vector
...Kozak 2 consensus sequence. Each ribosome binding site is positioned at its optimaldistance from the init...n addition, the thrombin protease cleavage site allows cleavage to occur in the presence of thrombin for applications where removal of the a...Mammalian Expression Vector for Efficient Cloning of PCR Fragments
... CMV promoter and the SV40 polyadenylation site allow high-level constitutive expression in a wide variety of cell lines. In addition, the neomycin-resistance gene is under control of the prokaryotic b -lactamase promoter to provide kanamycin resistance in bacteria as well...Mammalian Expression Vectors with Interchangeable Selectable Markers
...ences. 3,4,5 For Cre recombinase, the recognition site (LoxP) consists of two inverted 13-bp repeats and ...ors vectors ( Figure 1 ) that contain a LoxP site that serves as the acceptor site for the eukaryotic selectable-marker cassettes and offer a va...Sequence-Validated and Expression-Tested Human cDNA in a Dual Expression Vector
...e fusion protein also contains a thrombin cleavage site between GFP and the c-myc epitope. Followi...leotide Sequence Insertion The unique Not I site between the cDNA insert and thrombin cleavage site in the pDual GC vector allows any desired...New Fusion Trans-Activator Plasmids for Studying Signal Transduction Pathways
...in. The pFA-CMV plasmid features amultiple cloning site and the cytomegalovirus (CMV) promoter for conveni...MV plasmid features (1) a multiple cloning site (MCS) that offers 10 unique, conveniently arranged restriction sites for insertion of the tr...Cis-Reporting System Monitors Intracellular Calcium Mobilization
...a consensus sequence of T(A)GGAAAAT(A/C). 1,2 This site is present in many genes involved in immune respon...T-Luc reporterplasmid, containing the NFAT binding site from the human IL-2 gene. 2,7-9 The NFAT cis-reporting system includes the transfection-ready pNFAT-...Cis-Reporting System Monitors Intracellular Calcium Mobilization
...a consensus sequence of T(A)GGAAAAT(A/C). 1,2 This site is present in many genes involved in immune respon...T-Luc reporterplasmid, containing the NFAT binding site from the human IL-2 gene. 2,7-9 The NFAT cis-reporting system includes the transfection-ready pNFAT-...Decontamination and inhibition of ribonucleases
...zymes rely on histidine residues within the active site for catalytic activity and can be inactivated by the alkylating agent diethyl pyrocarbonate (DEPC) which modifies these residues. RNase contamination can effectively be avoided by using RNase-free solutions such as Eppendorf M...Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler
... -globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230: 1350-1354. (2) Persing, D.H. (1993). Target selection and optimization of amplification reaction. In: Diagnostic molecular microbiology, 88-102. Eds.: David H. ......of beta-globin sequences and restriction site analysis for diagnosis of sickle cell anemia." Science, 230: 1350-1354. Sharroks, A.D. 1994. "The design of primers for PCR," PCR Technology, Current Innovations, Griffin, H.G., and Griffin, A.M., Ed., CRC Press...TripleMaster PCR SystemThe Ideal Choice for Long-Range PCR
...aq Polymerase is unable to extend past the site of depurination and amplification of the desired target is not possible. Another crucial factor in the limitation of standard PCR to amplify long target sequences is the high rate of incorporation of incorrect bases by thermos...The QIAGEN Guide to Animal Cell Culture
...ded. Tip - The Transfection Tools web site (www.qiagen.com/transfectiontools/) has an online cell database that includes information about media used with different cell types. Click on the cell type of interest, and then click on Detail to view this information. ......y that a primer has more than one specific binding site within a genome is significantly lower for longer primers. In addition, longer primers allow annealing at slightly higher temperatures where Taq DNA polymerase activity is higher. GC cont...TransMessenger Transfection Reagent
...oducts for RNAi visit the Transfection Tools site at www.qiagen.com/transfectiontools ....Effectene Transfection Reagent
...tocols, is available at the Transfection Tools web site www.qiagen.com/transfectiontools/ . Serum and DNA Quantity vs. Transfection Efficiency Influence of serum and DNA quantity on...... Technical Services and the Transfection Tools web site ( www.qiagen.com/transfectiontools/ ). Poly... Services or visit the Transfection Tools web site at www.qiagen.com/transfectiontools/ . Cited References 1. Tang, M.X., Re...SuperFect Transfection Reagent
...is available at the Transfection Tools web site www.qiagen.com/transfectiontools/. Transfection of Neuronal PC-12 Cells Using SuperFect Reagent Expression of green fluorescent protein (GFP) in differentiated PC-12 cells 5 days post-transfection. 104105 c...Ni-NTA Superflow Columns - automated large-scale purification of 6xHis-tagged proteins
...3 chelating sites. The extra chelating site of NTA prevents nickel-ion leaching, increasing protein binding capacity and minimizing nonspecific binding. Automated procedure for large-scale purification Ready-to-run protocols for...TransMessenger Transfection Reagent Handbook
...as a cap, poly-A tail, or internal ribosomal entry site (IRES), may increase or decrease the efficie...on online The QIAGEN Transfection Tools web site contains siRNA transfection protocols and the most up-to-date information and literature on QI...TransMessenger Transfection Reagent
...oducts for RNAi visit the Transfection Tools site at www.qiagen.com/transfectiontools ....... this oxidation can vary depending on the specific site of the oxidation: MetSO at the Nterminal end...equencing these peptides and determining the exact site of oxidation. (This is especially useful for peptides containing Met residues) 3)...Structural Analysis of Glycosylated Peptides in Complex Mixtures with Ion Trap MSn
... the oligosaccharide structure nor the exact site of attachment to the peptide can be determined. Th... acid sequence of glycopeptides The exact site of attachment of the oligosaccharide Accurate, detailed structures of the attached oligosac...In-Vitro Metabolism Studies Using Data-Dependent LC/MSn
...ata obtained is not sufficient to locate the site of metabolism on a candidate molecule. The Thermo ... experiments to locate more specifically the site of metabolism. Since the samples utilized microsomal preparations fortified with NADPH, only o......gment structure that does contain the substitution site (i.e. if the fragment ion has a m/z value of...ss Frontier prediction contains the proposed site of substitution. This observation provides the main verification for the impurity structures t......is of MS/MS data reveals the serine residue as the site of phosphorylation (Figure 6). Figure 5. Neurofilament protein identified in TurboSEQUEST search. The peptide (in red) is phosphorylated at its serine residue (highlighted in green). ......ompilation Data delivery via a secure FTP site COMPREHENSIVE AND PROVEN PIPELINE Agencourts comprehensive pipeline allows for effortless transition from end sequencing to complete insert sequence delineation. We have extensive experience with both end ......nd finishing Data delivery via a secure FTP site FULLY-AUTOMATED, GLP-COMPLIANT FACILITY Agencourt has a fully-automated genomic services pipeline based on SPRI, our patented nucleic acid purification technology. All aspects of our pipeline are managed by......alysis Data delivery via secure FTP site ASSAY DESIGN AND VALIDATION A notable strength of Agencourt is our extensive assay design and validation process. A suite of proprietary software tools is used to design amplification primers and develop ...... 3700/3730xl Data delivery via a secure FTP site HIGH QUALITY SEQUENCING RESULTS Our fully-automated genomic pipeline and SPRI nucleic acid purification technology allow us to process over 20 million Phred20 bases per day. We use this large through......ompilation Data delivery via a secure FTP site UNCOMPROMISED DATA & SAMPLE INTEGRITY Agencourts Oracle-based LIMS continually monitors sample genealogy and quality parameters from sample submission through data delivery. In ...A Further Step in Understanding Apoptosis Direct Detection of PARP Cleavage
...ssince it is directed against the caspase cleavage site ofPARP-1 and specifically recognizes an epitope that isgenerated by caspases, but is not accessible in nonapoptoticcells. This easy-to-use addition to the Roche caspase-signalingproduct line provides a number of benefits, including Fast,......or anymutant sequences. In mutatedsamples, a BstNI site was notcreated, and therefore not recognized.Tumor...d. Ifthe sample was wild-type, BstNIrecognized its site and trimmedthe product to a 106 bp size. If thesample was mutated, the digestedproduct remained at 1...