The pd1pdl1 blockade bioassay includes the necessary medium and serum to thaw, plate and assay the cells. The iscript reverse transcription supermix for rtqpcr reaction setup for a single cdna synthesis reaction iscript rt supermix is a sensitive, fast, and convenient reagent for gene expression analysis using realtime reverse transcription quantitative pcr rtqpcr and standard rtpcr. Instructions for use of products j1250, j1255, j3011, j3015, j4011, j4015. Moloney murine leukemia virus reverse transcriptase, rnase h minus, point mutant mmlv rt h is an rnadependent dna polymerase that can be used in cdna synthesis with long messenger rna templates 5kb. Monarch nucleic acid purification kits are optimized for maximum performance and minimal environmental impact. The adpglo kinase assay is a luminescent adp detection assay that provides a universal, homogeneous, highthroughput screening method to measure kinase activity by quantifying the amount of adp produced during a kinase reaction. The chemistry eliminates tedious centrifugation steps as well as the use of hazardous chemicals, which are inherent in precipitationbased chemistries. Product components and storage conditions product size cat. Free isotope was removed by sephadex g50 column chromatography pharmacia, uppsala, sweden. Mmlv rt is an rnadependent dna polymerase that can be used in cdna synthesis with.
Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. New england biolabs is working diligently to ensure we keep our employees and their families safe, while maintaining our business continuity. The pd1pdl1 blockade bioassay is a bioluminescent cellbased assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the pd1pdl1 interaction. In many cases, good activity is also obtained using one of the. Following incubation, the supercoiled dna is visualized on an ethidium bromidestained agarose gel to verify the absence of visible nicking or cutting analysis on 0. Mar 12, 2009 unauthorized use andor duplication of this material without express and written permission from this promega corporation is strictly prohibited. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. Reliaprep large volume ht gdna isolation system protocol. Print anything, with two cubic feet of build space. Molecular weight markers, or ladders, are a set of standards that are used for determining the approximate size of a protein or a nucleic acid fragment run on an electrophoresis gel. For maximum convenience and value, columns and buffers are also available separately.
Product source the gene encoding mmulv reverse transcriptase is expressed in li in a vector that results in 16 additional amino acids at the nterminus and amino acids at the cterminus. Superscript iii rt offers higher cdna yields, improved cdna lengths, improved efficiency on gcrich target rnas, and overall better performance than wildtype mmlv and mmlv rnase hminus enzymes. Moloney murine leukemia virus reverse transcriptase mmlv rt uses single stranded rna or dna in the presence of a primer to synthesize. Both probe and dyebased systems provide sensitive detection for reproducible and earlier quantification of low and highcopynumber targets over a broad dynamic range, together with resistance to a wide range of pcr inhibitors. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide. Despite a stay at home advisory being put in place in massachusetts, usa, we are deemed an essential business, and our manufacturing and distribution teams continue to be fully operational. The gene encoding mmulv reverse transcriptase is expressed in li in a vector that results in 16 additional amino acids at the nterminus and amino acids at the cterminus. Three sizes of kit are available 10, 50 and 250 reactions and each kit contains membrane binding solution, membrane wash solution, nucleasefree water, a set. Avian myeloblastosis virus amv reverse transcriptase is an rnadirected dna polymerase. T4 dna ligase bluewhite cloning qualified protocol promega. Mmlv rt is an rnadependent dna polymerase that can be used in cdna synthesis with long messenger rna templates 5kb.
The enzyme is provided preblended with rnase inhibitor. Instructions for use of products e2920, e2940, e2980. This construct results in a fully functional reverse transcriptase protein with a functional rnase h domain. The dualglo luciferase assay system is designed to allow highthroughput analysis of mammalian cells containing genes for firefly and renilla luciferases, grown in 96 or 384well plates. Fulllength cdna is observed by gel electrophoresis and autoradiography. Novel mutations in moloney murine leukemia virus reverse. Mmlv reverse transcriptase, rnase h minus, point mutant.
The activity of t4 polymerase and mmlv and amv reverse transcriptase. Picomaxx dna polymerase blend of taq and pfu, human total rna skeletal muscle, hela, human universal reference total rna, genemorph ii ezclone domain mutagenesis kit, strataprep plasmid miniprep kit, quikchange sitedirected mutagenesis kit, quikchange multi sitedirected mutagenesis kit, and bl21codonplus de3ril competent cells were from stratagene. Promega corpora on 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 3 tm432 315 3. Refer to the agilent oligonucleotide arraybased cgh for genomic dna analysis enzymatic labeling for blood, cells, or tissues protocol pn g441090010.
Promega mmlv reverse transcriptase mmlv rt is an rnadependent dna polymerase that can be used in cdna synthesis with long messenger rna templates 5kb. Most men with lowgrade confined primary pca are generally curable by surgery and radiotherapy 2,3. Developmental validation of the powerplex r 18d system, a rapid str multiplex for analysis of reference samples. Prostate cancer pca is the second commonly diagnosed cancer in men worldwide 1. Amv reverse transcriptase, mmlv reverse transcriptase. Ethanol down regulates the expression of myelin proteolipid.
All publications and patents mentioned in the above specification are herein. Dna quantitation in nextgeneration sequencing library workflows. The adpglo kinase assay is performed in a multiwell plate and can detect kinase activity in a reaction volume as low as 5. Excerpts and links may be used, provided that full and clear credit is given to promega corporation with appropriate and specific direction to the original content. Moloney murine leukemia virusmmlv reverse transcriptase is an rnadependent. Nov 08, 2006 the wizard sv gel and pcr cleanup system is a multipurpose kit that allows fragments of dna to be extracted from gel slices or directly from small volume pcr or enzymatic reactions.
Us6242235b1 polymerase stabilization by polyethoxylated amine. These standards contain predetermined fragment or protein sizes and concentrations. Sv lysis buffer use in sv and sv 96 genomic dna purification system. Restriction enzyme activity in promega 10x buffers. This enzyme can synthesize a complementary dna strand initiating from a primer using rna cdna synthesis or singlestranded dna as a template. The amount of input dna depends on the ngs library preparation method, as well as the specific type of ngs application. The cell lines listed in table 1 have been successfully transfected by promega corporation or fugent, l. Instructions for use of products v6930, v9101, v9102, v9103, v9104.
Mmlv reverse transcriptase in 1x reaction buffer for one hour at 37c. The bioluminescent signal can be detected and quantified using the bioglo luciferase assay system, also included in the kit, and a standard luminometer such as the glomax discover system. Posted on march 12, 2009 june 17, 2014 by kelly grooms. Labelled probes were denatured by boiling for 10 min and. See our terms and conditions for additional information. Mar 19, 2012 unauthorized use andor duplication of this material without express and written permission from this promega corporation is strictly prohibited.
Promega real time pcr qpcr and rtqpcr systems provide a choice of probebased or dyebased qpcr methods. The vector is engineered with fewer consensus regulatory sequences for reduced backgrounds and a decreased risk of anomalous transcription. Mmlv reverse transcriptase certificate of analysis. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences. The promega is reliable, fast and overengineered to ensure high quality printing. Mmlv reverse transcriptase thermo fisher scientific. The minimum specification is 120ng of firststrand cdna made from 1g of. Dualluciferase reporter assay system protocol promega. Agilent oligonucleotide arraybased cgh for genomic dna.
The enzyme has also been shown to catalyze the joining of rna to either a dna or rna strand in a duplex molecule, but will not join singlestranded. The 10x reaction buffer supplied with each restriction enzyme is optimized to give 100% activity. Product source insect cells sf21 infected with baculovirus containing the pol gene of amv. Mmlv reverse transcriptase protocolpdf 110 kb english. Rnasin ribonuclease inhibitor product informationpdf promega.
Polymerase stabilization by polyethoxylated amine surfactants download pdf. T4 dna ligase catalyzes the joining of two strands of dna between the 5. Pdf developmental validation of the powerplex r 18d. Restriction enzyme activity in promega 10x buffers, reaction temperature and heat inactivation. The reliaprep large volume ht gdna isolation system isolates genomic dna from 1ml to 10ml of blood in a scalable format. The cdna product must be 90% full length as determined by gel electrophoresis.
Promega corporation 2800 woods hollow rd, madison, wisconsin 53711 rated 4. Promega qualified and verified the rnasin ribonuclease inhibitor manufacturing process for its viral. However, the clinical challenge is to provide effective means for treatment of patients whose cancer advances to a deadly, latestage, metastatic pca. Nen, 200 units of mmlv reverse transcriptase promega, fitchburg, wisconsin and 20 units of rnasin were added and incubated at 37oc for 1 hr. Agilent oligonucleotide arraybased cgh for genomic dna analysis.