For focused group profiling of human cell cycle toxicity and cancer related gene expression

Transcription

1 GeneCopoeia TM Expressway to Discovery Product Data Sheet ExProfile TM Human Cell Cycle Toxicity and Cancer Related Gene qpcr Array For focused group profiling of human cell cycle toxicity and cancer related gene expression Cat. No. QG053-A (1 x 96-well plate, Format A) Cat. No. QG053-B (1 x 96-well plate, Format B) Cat. No. QG053-C (1 x 96-well plate, Format C) Cat. No. QG053-D (1 x 96-well plate, Format D) Cat. No. QG053-E (1 x 96-well plate, Format E) Plates available individually or as a set of 6. Each set contains 84 unique gene primer pairs deposited in one 96-well plate. Introduction The ExProfile human cell cycle toxicity and cancer related gene qpcr array profiles the expression of 84 cell cycle genes important in both toxicology and cancer research. These genes are carefully chosen for their close cell cycle correlation based on a thorough literature search of peer-reviewed publications, and include genes that encode cyclins, cyclin-dependent kinases and phosphatases, cell cycle inhibitors, and genes important for DNA replication, cycle checkpoints, and cell cycle arrest. This array allows researchers to study pathway-related genes to gain understanding of their roles during cancer or drug treatment. QG053 plate 01: 84 unique gene PCR primer pairs Shipping and storage conditions Shipped at room temperature Stable for at least 6 months when stored at -20 C Array format GeneCopoeia provides five qpcr array formats (A, B, C, D, and E) suitable for use with the following realtime cyclers. Important note: Upon receipt, please check to make sure that the correct array format was ordered to ensure compatibility with your qpcr instrument. Plate format Instrument provider qpcr instrument model A B C D E Applied Biosystems Applied Biosystems Bio-Rad Laboratories Bio-Rad Laboratories Roche Applied Science 5700, 7000, 7300, 7500, 7700, 7900HT (Standard 96-well block), ViiA TM 7 (Standard 96-well block) 7500 (Fast block), 7900HT (Fast block), StepOnePlus TM, ViiA TM 7(Fastblock) icycler iq,myiq,iq 5 CFX96, DNA Engine Opticon, DNA Engine Opticon 2, Chromo4 LightCycler 480 (96-well block) 1

2 Quality control 1. Each pair of primers in the ExProfile gene qpcr array has been experimentally validated to yield a single dissociation curve peak and to generate a single amplicon of the correct size for the targeted gene. 2. The positive PCR controls (PCR) have been verified to amplify a single amplicon of the correct size with Ct values around 20±2. 3. The Spike-in reverse transcription controls (RT) have been verified to amplify a single amplicon of the correct size with Ct values around 20±3. 4. R 2 > 0.99 was observed for high inter/ intra-array reproducibility. Materials required but not provided All-in-One TM First-Strand cdna Synthesis Kit All-in-One TM qpcr Mix Total RNA extraction kit (RNAzol RT RNA extraction reagent is recommended) DNase/RNase free tips, PCR reaction tubes, 1.5 ml microcentrifuge tubes 5 ml and 10 ml graduated pipettes, beakers, flasks, and cylinders 10 μl to1,000μl adjustable single channel micropipettes with disposable tips 5 μl to20μl adjustable multichannel micropipette, disposable tips, and reservoir qpcr instrument, compatible with gene qpcr arrays ordered Array layout Figure1. Illustration of QG053 plate 01 Gene primer pairs: 84 wells (A row to G row) are designated for a real-time PCR assay for genes (see the primer list). HK1-6: Six pre-deposited housekeeping gene (HK1-6) primer pairs, which can be used as endogenous positive controls as well as for array normalization. GDC: Genomic DNA controls, which can be used to specifically detect genomic DNA contamination with a high level of sensitivity. RT: Spike-in reverse transcription controls, which can be used to monitor the efficiency of the RT reactions. These pre-deposited primer pairs specifically amplify the cdna template reverse transcribed from the spike-in control RNA in the sample. PCR: Positive PCR controls, which are used to verify the PCR efficiency by amplifying the predeposited DNA template with its specific pre-deposited primer pairs. 2

3 Gene primer list Plate Position Catalog No. of Primer Accession No. of Gene Symbol QG A01 HQP NM_ ABL1 QG A02 HQP NM_ AHR QG A03 HQP NM_ AKT2 QG A04 HQP NM_ ANAPC10 QG A05 HQP NM_ ATM QG A06 HQP NM_ BRCA1 QG A07 HQP NM_ CCNA2 QG A08 HQP NM_ CCNB1 QG A09 HQP NM_ CCNB2 QG A10 HQP NM_ CCND1 QG A11 HQP NM_ CCND2 QG A12 HQP NM_ CCND3 QG B01 HQP NM_ CCNE1 QG B02 HQP NM_ CCNG1 QG B03 HQP NM_ CCNG2 QG B04 HQP NM_ CCNH QG B05 HQP NM_ CCNI QG B06 HQP NM_ CCNK QG B07 HQP NM_ CDC14A QG B08 HQP NM_ CDC16 QG B09 HQP NM_ CDC2 QG B10 HQP NM_ CDC20 QG B11 HQP NM_ CDC23 QG B12 HQP NM_ CDC25A QG C01 HQP NM_ CDC25B QG C02 HQP NM_ CDC25C QG C03 HQP NM_ CDC27 QG C04 HQP NM_ CDC2L5 QG C05 HQP NM_ CDC37 QG C06 HQP NM_ CDC45L QG C07 HQP NM_ CDC6 QG C08 HQP NM_ CDK2 QG C09 HQP NM_ CDK4 QG C10 HQP NM_ CDK6 QG C11 HQP NM_ CDK7 QG C12 HQP NM_ CDK9 QG D01 HQP NM_ CDKL1 QG D02 HQP NM_ CDKN1A QG D03 HQP NM_ CDKN1B QG D04 HQP NM_ CDKN2A QG D05 HQP NM_ CDKN2C 3

4 4 Product Data Sheet QG D06 HQP NM_ CDKN2D QG D07 HQP NM_ CDKN3 QG D08 HQP NM_ CETN2 QG D09 HQP NM_ CHEK1 QG D10 HQP NM_ CHEK2 QG D11 HQP NM_ CUL1 QG D12 HQP NM_ DDIT3 QG E01 HQP NM_ DMC1 QG E02 HQP NM_ DUSP1 QG E03 HQP NM_ E2F1 QG E04 HQP NM_ E2F2 QG E05 HQP NM_ E2F3 QG E06 HQP NM_ E2F4 QG E07 HQP NM_ GADD45A QG E08 HQP NM_ HDAC1 QG E09 HQP NM_ HRAS QG E10 HQP NM_ JUN QG E11 HQP NM_ JUNB QG E12 HQP NM_ MAD2L2 QG F01 HQP NM_ MAPK1 QG F02 HQP NM_ MCM2 QG F03 HQP NM_ MCM3 QG F04 HQP NM_ MCM6 QG F05 HQP NM_ MDM2 QG F06 HQP NM_ MPHOSPH1 QG F07 HQP NM_ MYC QG F08 HQP NM_ NRAS QG F09 HQP NM_ POLS QG F10 HQP NM_ PTEN QG F11 HQP NM_ RAD17 QG F12 HQP NM_ RAD21 QG G01 HQP NM_ RAF1 QG G02 HQP NM_ RB1 QG G03 HQP NM_ TAF10 QG G04 HQP NM_ TFDP1 QG G05 HQP NM_ TFDP2 QG G06 HQP NM_ TP53 QG G07 HQP NM_ UHRF1 QG G08 HQP NM_ WEE1 QG G09 HQP NM_ WT1 QG G10 HQP NM_ FOSB QG G11 HQP NM_ ORC1L QG G12 HQP NM_ PKMYT1 QG H01 HGDC QG H02 HGDC QG H03 HQP NM_ GAPDH QG H04 HQP NM_ ACTB

5 QG H05 HQP NM_ B2M QG H06 HQP NM_ RPL13A QG H07 HQP NM_ HPRT1 QG H08 HQP NR_ RN18S1 QG H09 RT QG H10 RT QG H11 PCR QG H12 PCR 5

6 Limited Use License Following terms and conditions apply to use of ExProfile TM Human Cell Cycle Toxicity and Cancer Related Gene qpcr Array (the Product). If the terms and conditions are not acceptable, the Product in its entirety must be returned to GeneCopoeia within 5 calendar days. A limited End-User license is granted to the purchaser of the Product. The Product shall be used by the purchaser for internal research purposes only. The Product is expressly not designed, intended, or warranted for use in humans or for therapeutic or diagnostic use. The Product must not be resold, repackaged or modified for resale, or used to manufacture commercial products or deliver information obtained in service without prior written consent from GeneCopoeia. This Product should be used in accordance with the NIH guidelines developed for recombinant DNA and genetic research. Use of any part of the Product constitutes acceptance of the above terms. Limited Warranty GeneCopoeia warrants that the Product meets the specifications described in the accompanying Product Datasheet. If it is proven to the satisfaction of GeneCopoeia that the Product fails to meet these specifications, GeneCopoeia will replace the Product. In the event a replacement cannot be provided, GeneCopoeia will provide the purchaser with a refund. This limited warranty shall not extend to anyone other than the original purchaser of the Product. Notice of nonconforming products must be made to GeneCopoeia within 30 days of receipt of the Product. GeneCopoeia s liability is expressly limited to replacement of Product or a refund limited to the actual purchase price. GeneCopoeia s liability does not extend to any damages arising from use or improper use of the Product, or losses associated with the use of additional materials or reagents. This limited warranty is the sole and exclusive warranty. GeneCopoeia does not provide any other warranties of any kind, expressed or implied, including the merchantability or fitness of the Product for a particular purpose. GeneCopoeia is committed to providing our customers with high-quality products. If you should have any questions or concerns about any GeneCopoeia products, please contact us at GeneCopoeia, Inc. GeneCopoeia, Inc Medical Center Drive, Suite 101 Rockville, MD (301) (866) inquiry@genecopoeia.com GeneCopoeia Products are for Research Use Only Copyright 2016 GeneCopoeia, Inc. Trademarks: GeneCopoeia, ExProfile, All-in-One, (GeneCopoeia Inc.); RNAzol RT (Molecular Research Center, Inc.); SYBR (Molecular Probes); icycler iq, MyiQ, iq 5, CFX96, DNA Engine Opticon, DNA Engine Opticon 2, Chromo4 (Bio-Rad); LightCycler (Roche);Trizol, ABI,ROX,ViiA TM,StepOnePlus TM (Life Technologies). QG