Science Catalog 2012 Life Cell Signaling Cell Line Authentication StemElite™ ID System Product StemElite™ ID System Available Separately Water, Amplification Grade 9947A DNA Internal Lane Standard 600 Size Conc. 6,250 µl Size Cat.# Price (Fr) 50 reactions G9530 1676.00 Cat.# Price (Fr) DW0991 47.00 250 ng 10 ng/µl DD1001 Pls. Enq. 150 µl DG1071 472.00 DW0991, DG1071 For Laboratory Use. G9530 For Research Use Only. Not for Use in Diagnostic Procedures. DD1001 Not For Medical Diagnostic Use. Description: The StemElite™ ID System provides the reagents required for successful and simple identification and authentication of human cell lines and detection of intra-species cell line cross-contamination. The StemElite™ ID System uses short tandem repeat (STR) analysis of specific loci in the human genome through co-amplification and three-color detection of ten loci (nine STR loci and Amelogenin for gender identification), including D21S11, TH01, TPOX, vWA, Amelogenin, CSF1PO, D16S539, D7S820, D13S317 and D5S818. Additionally, the StemElite™ ID System incorporates a sensitive marker that specifically detects the presence of DNA from mouse (Mus musculus). All eleven loci are amplified simultaneously in a single tube. The StemElite™ ID System includes a hot-start Taq DNA polymerase in an enzyme mix for convenient room-temperature reaction assembly. Alleles, along with the allelic ladder and an internal lane standard that assists in assigning sizes for each of the loci, are resolved by capillary electrophoresis in a single injection, and CE data are analyzed using allele-calling software to generate a genetic profile of the cell line. These loci collectively provide a genetic profile with a random match probability of 1 in 2.92 × 109. The StemElite™ ID System is designed specifically for use with the Applied Biosystems 3130 and 3130xl Genetic Analyzers, ABI PRISM® 3100 and 3100-Avant Genetic Analyzers and ABI PRISM® 310 Genetic Analyzer. Dye matrix generation is required on initial setup and is performed using the PowerPlex® Matrix Standards, 310 (Cat.# DG4640), or PowerPlex® Matrix Standards, 3100/3130 (Cat.# DG4650). Features: • Improved “Gold Standard” STR Analysis: The StemElite™ ID System includes all loci used in the original PowerPlex® 1.2 System plus the D21S11 locus for improved power of discrimination. • Detects Mouse Contamination: The StemElite™ ID System includes a mouse locus, allowing detection of contamination of human cells with mouse cells. • Simplified Data Interpretation: The StemElite™ ID System allows co-amplification of 10 loci to accurately discriminate between human cell lines without using a full forensic profile and includes allelic ladders for all 10 loci to simplify data interpretation. • Simplified Amplification Setup: Inclusion of a hot-start Taq DNA polymerase allows room-temperature reaction assembly. • Internal Lane Standard 600 Included: This marker offers the greatest precision available for DNA typing. It is designed for use in each capillary injection to increase the precision of your analyses. • Automatic Assignment of Genotypes: To make genotyping easier and more accurate, panels and bins text files are available for download and are required for use with GeneMapper® ID software. GeneMapper® 4.0 software requires custom bin generation. Storage Conditions: Store at –20°C. The Primer Pair Mix, Allelic Ladder and Internal Lane Standard are light-sensitive and should be stored protected from light. Protocol StemElite™ ID System Technical Manual Fluorescein: JOE: TMR: CXR: Size (bp) M D5S818 A vWA TH01 D21S11 D13S317 D7S820 D16S539 TPOX CSF1PO Part# TM307 100 A = Amelogenin M = Mouse Allele ranges for the StemElite™ ID System. STR fragments amplified by the StemElite™ ID System are labeled with different dyes and are separated by capillary electrophoresis based on size. A size standard is included in each sample to determine the sizes of the StemElite™ ID-amplified fragments. Fluorescein-labeled loci are shown in dark gray, JOE-labeled loci are shown in gray and TMR-labeled loci are shown in white. The CXR-labeled Internal Lane Standard 600 fragments are represented by black bars. 200 300 400 Representative data for the StemElite™ ID System. Genomic DNA from 9947A cells (2.0ng) combined with 20pg of mouse genomic DNA prior to amplification was amplified using the StemElite™ ID System. Amplification products were mixed with Internal Lane Standard 600 and analyzed with an Applied Biosystems 3130xl Genetic Analyzer using a 1.5kV, 5-second injection. The results were analyzed using GeneMapper® ID software, version 3.2, and the appropriate panels and bins text files. 294 For complete and up-to-date product information visit: www.promega.com/catalog 7818TA 7815MA Pagina 297
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