Package Insert, GeneAtlas Hyb, Wash, and Stain Kit for WT Array
Contents
1. by Affymetrix is conveyed or implied by this Affymetrix product In particular no right or license is conveyed or implied to use this Affymetrix product in combination with a product not provided licensed or specifically recommended by Affymetrix for such use Trademarks Affymetrix Axiom Command Console DMET GeneAtlas GeneChip GeneChip compatible GeneTitan Genotyping Console NetAffx and Powered by Affymetrix are trademarks or registered trademarks of Affymetrix Inc All other trademarks are the property of their respective owners Copyright 2010 Affymetrix Inc All rights reserved P N 702939 Rev 1 Ct CK Keference CGttde Affymeini Hybridization Cocktail Preparation Instructions for Processing Gene 1 1 ST Array Strips using the GeneAtlas Instrument and the WT Assay The Hybridization Mix described in this Quick Reference Guide was specifically formulated for use Note The WT Hyb Add reagent names with the Ambion WT Expression Kit and the GeneChip WT Terminal Labeling and Controls Kit were created to match the order inaahich reagents are added For example WT Hyb 1 Remove the vials labeled 5X WT Hyb Add 1 15X WT Hyb Add 4 and 2 5X WT Hyb Add 6 Add 4 is the fourth component added during from the GeneAtlas Hybridization Module for WT Array Strips P N 901668 preparation of the Hybridization Mix WT Hyb Add 2 3 and 5 are not used and are not part A Warm reagents to room tempe2. KG Package Affymetrix Insert GeneAtlas Hybridization Wash and Stain Kit for WT Array Strips 60 Reactions Intended Use This package insert provides information for the GeneAtlas Hybridization Wash and Stain Kit for WT Array Strips P N 901667 The kit is composed of two boxes m P N 901668 GeneAtlas Hybridization Module for WT Array Strips sufficient for 60 reactions m P N 901625 GeneAtlas Wash Buffers A and B Module sufficient for 60 array Strips The GeneAtlas Hybridization Wash and Stain Kit for WT Array Strips includes all the necessary reagents for the hybridization wash and stain steps of the Ambion WT Expression Kit and GeneChip WT Terminal Labeling and Controls Kit using Atfymetrix Whole Transcript WT Array Strips Module Components Component Volume Storage GeneAtlas Hybridization Module for WT Array Strips P N 901668 5X WT Hyb Add 1 P N 901618 3 x925 uL 2 C to 8 C 15X WT Hyb Add 4 P N 901619 3 X 359 UL 2 C 10 8C 2 5X WT Hyb Add 6 P N 901620 3 x 1 525 uL 2 C to 8 C Stain Cocktail 1 P N 901541 52 5 Tne IAG 1O EC Stain Cocktail 2 P N 901542 26 3 mL 2 C to 8 C Array Holding Buffer P N 901543 15 mL 2 C TO 8PC GeneAtlas Wash Buffers A amp B Module P N 901625 Wash Buffer A 5 9 ime 2 C TO 8PC Wash Buffer B 64 mL 2 C to 8 C Note The WT Hyb Add reagent names were created to match the order in which reagents are added For example WT Hyb Add 4
3. is the fourth component added during preparation of the Hybridization Mix WT Hyb Add 2 3 and 5 are not used and are not part of the Hybridization Modules Shipping Information The components of the GeneAtlas Hybridization Wash and Stain Kit are shipped at different temperatures The GeneAtlas Hybridization Module for WT Array Strips P N 901668 will be sent at 2 C to 8 C The GeneAtlas Wash Buffers A amp B Module P N 901625 will be sent at ambient temperature Safety Information Warning P N 901668 contains Tetramethylammonium chloride TMACI and formamide both of which are considered highly toxic P N 901668 contains Sodium Azide May react with lead and copper plumbing to form highly explosive metal azides A Material Safety Data Sheet s MSDS is available at www affymetrix com If the product is a kit or is supplied with more than one material please refer to the MSDS for each component for hazard information Functional Testing Each lot of reagents is rigorously tested for performance GeneAtlas Hybridization Wash and Stain Kit for WT Array Strips For Research Use Only Not for use in diagnostic procedures Ordering Information P N Product Name Description Size 901667 GeneAtlas Hybridization Wash and Stain Kit 60 Rxn for WT Array Strips Related Products 4411973 Ambion WT Expression Kit 10 Rxns 900670 GeneChip WT Terminal Labeling Kit 10 Rxns Affymetrix products can be purchased directly f
4. ntration Reagents Array 5 Fragmented and Labeled DNA 41 uL 25 ng uL Total Volume 90 uL C Add the 2 5X WT Hyb Add 6 from the GeneAtlas Hybridization Module for WT Array Strips as shown in Table 3 Table 3 Order to Add Component Volume per Final Concentration Reagents Array 6 2 5X WT Hyb Add 6 60 uL 1X Total Volume 150 uL D If you are using a plate seal vortex and centrifuge briefly 5 sec to collect liquid at the bottom of the well If you are using 1 5 mL tubes vortex and centrifuge briefly 5 sec to collect liquid at the bottom of the tube 4 Denature the hybridization cocktail with target at 99 C 1 5 mL tubes or 95 C thermocycler plates for 5 minutes followed by 45 C for 5 minutes 5 After denaturation spin hybridization cocktail with target in a centrifuge to remove any insoluble material from the hybridization mixture If you are using 1 5 mL tubes use an Eppendorf 5417C centrifuge or equivalent If you are using thermocycler plates use an Eppendorf 5804R centrifuge or equivalent Spin tubes or plates for 1 minute at 5000 RPM at room temperature 6 Place 120 uL of the centrifuged supernatant hybridization cocktail with target into the appropriate well of the hybridization tray 7 Refer to the GeneAtlas System User s Guide P N 08 0306 for details on the GeneAtlas hybridization setup For Research Use Only Not for use in Diagnostic Procedures
5. rature on the bench B Vortex the 5X WT Hyb Add 1 15X WT Hyb Add 4 and 2 5X WT Hyb Add 6 to mix Centrifuge briefly 5 sec to collect liquid at the bottom of the tube of the Hybridization Modules 2 Remove the GeneChip Hybridization Control Kit from 20 C freezer and thaw at room temperature A Vortex and centrifuge briefly 5 sec to collect liquid at the bottom of the tube B Keep on ice 3 WT Hybridization Master Mix A new hybridization mix has been formulated for the Ambion WT Expression Kit and the GeneChip WT Terminal Labeling and Controls Kit A Prepare the WT Hybridization Master Mix in the order as shown in Table 1 The 5X WT Hyb Add 1 solution is very viscous pipet slowly to ensure addition of the correct volume Mix well Table 1 Order to Add Component Volume per Array 4 Array Strip Final Concentration Reagents 1 5X WT Hyb Add 1 30 uL 132 uL 1X 2 Control Oligonucleotide B2 3 nM 1 8 wl 6 6 uL 30 pM 3 20X Eukaryotic Hybridization Controls 7 5 UL 33 uL 1 5 5 25 and 100 pM bioB bioC bioD cre respectively 4 15X WT Hyb Add 4 10 ul 44 uL 1X Total Volume 49 uL 215 6 uL Includes 10 overage to cover pipetting error B Aliquot 49 uL of the master mix prepared in Table 1 to each tube or well Add the fragmented and labeled single stranded DNA target generated from the GeneAtlas WT Expression Kit User Manual P N 702935 as shown in Table 2 Table 2 Order to Add Component Volume per Final Conce
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