Cell Signaling HeLaScribe® Nuclear Extract in vitro Transcription System Product HeLaScribe® Nuclear Extract in vitro Transcription System For Research Use Only. Not for Use in Diagnostic Procedures. Description: The most well characterized cell-free system for in vitro transcription of eukaryotic genes is derived from HeLa cell nuclei. HeLa nuclear extracts can support accurate transcription initiation by RNA polymerase II and exhibit both basal and regulated patterns of RNA polymerase transcription. The nuclear extract is also a source for a variety of transcription factors, DNAbinding proteins and the enzymatic machinery involved in RNA processing. The HeLa Nuclear Extract included in the HeLaScribe® Nuclear Extract in vitro Transcription System is prepared by a modification of the method of Dignam et al. Extracts prepared by this method have been shown to allow transcription from the human transferrin gene promoter and the adenovirus 2 major late promoter. The system also includes all of the necessary components for in vitro transcription as well as a positive control template (CMV immediate early promoter DNA). Features: • Peformance-Tested: Tested with cytomegalovirus immediate early gene (CMV) promoter. • Convenient: Available as a complete transcription system or extract alone. • Positive Control: System contains a CMV promoter-positive control template. Storage Conditions: Store at –70°C. Avoid multiple freeze-thaw cycles of the extract. Protocol HeLaScribe® Nuclear Extract in vitro Transcription System Technical Bulletin In vitro Transcription Systems Related Products Product HeLaScribe® Nuclear Extract in vitro Transcription Grade HeLaScribe® Nuclear Extract Positive Control DNA rCTP, rATP, rUTP, rGTP, 100mM each rATP, 100mM rUTP, 100mM rGTP, 100mM rCTP, 100mM Size Cat.# Price (Fr) 40 reactions E3091 426.00 160 reactions E3092 1364.00 300 ng E3621 106.00 4 × 400 µl E6000 493.00 400 µl E6011 154.00 400 µl E6021 154.00 400 µl E6031 154.00 400 µl E6041 154.00 E6000, E6011, E6021, E6031, E6041 For Laboratory Use. E3091, E3092, E3621 For Research Use Only. Not for Use in Diagnostic Procedures. Description: HeLaScribe® Nuclear Extract, in vitro Transcription Grade, derived from HeLa cell nuclei, provides a cell-free system for in vitro transcription of eukaryotic genes. Storage Conditions: Store HeLaScribe® Nuclear Extracts at –70°C. Store other components at –20°C. Protocol HeLaScribe® Nuclear Extract in vitro Transcription System Technical Bulletin Part# TB123 Part# TB123 M bp 311 – 249 – 200 – 151 – 140 – 118 – 100 – 82 – 66 – 48 – 42 – 40 – Size Cat.# Price (Fr) 40 reactions E3110 578.00 Primer Extension System—AMV Reverse Transcriptase Product Primer Extension System—AMV Reverse Transcriptase For Research Use Only. Not for Use in Diagnostic Procedures. Description: Primer Extension System—AMV Reverse Transcriptase can be used to quantitate specific mRNA transcripts and map the start sites of transcription. An end-labeled oligonucleotide is hybridized to RNA and is used as a primer by reverse transcriptase in the presence of deoxynucleotides. The RNA is thus reverse transcribed into cDNA and is analyzed on a denaturing polyacrylamide gel. The length of the cDNA reflects the number of bases between the labeled nucleotide of the primer and the 5´-end of the RNA; the quantity of cDNA product is related to the amount of targeted RNA. Features: • Convenient: System includes control RNA and primer as well as size markers ready for phosphorylation with T4 Polynucleotide Kinase. Storage Conditions: All components must be stored at –20°C, except for the control RNA, which must be stored at –70°C. Protocol Primer Extension System—AMV Reverse Transcriptase Technical Bulletin 0 10 5 2 ng control RNA Part# TB113 Size Cat.# Price (Fr) 40 reactions E3030 278.00 16 24 – Gel analysis of 32P-labeled ΦX174 DNA/HinfI markers and control RNA primer extension products produced using the Primer Extension System— AMV Reverse Trancriptase (Cat.# E3030). For complete and up-to-date product information visit: www.promega.com/catalog 289 RNA Analysis 0055TA01_6A Pagina 292
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