Bio-rad Aurum™ Total RNA 96 Kit Manual do Utilizador

Catalog #732-6800Aurum™ Total RNA 96 Kit Instruction Manual

6 Disruption and Homogenization Proper disruption and homogenization of the starting materials are required to ensure complete lysis of the cells an

7 Preparing the Aurum™ Vacuum Manifold Tubing provided in the Aurum™ Vacuum Manifold kit is 4 ft long and must be cut into appropriate pieces before

8 2. Place the desired 96-well binding plate on the manifold top and apply the recommended vacuum pressure for your application. Manifold Elution

9 Section 7 Vacuum Protocol PleasereadSection5,“BeforeUsingtheAurum™TotalRNA96Kit”andSection 6, “Vacuum Manifold Setup and Use With 96-

10 Yeast Follow steps C1–C5, then continue with step 1 of “All Starting Cell Types” onpage10.Ifstartingwithagrowblockofyeastculture(max

11 5. Add700µloflowstringencywashsolutiontoeachwelloftheRNAbinding plate. Gradually increase the negative pressure between –17 to –23

12 Note: Gradual application of negative pressure is required to prevent sample spraying and cross-contamination. The eluted total RNA sample

13B2. Add350µloflysissolution(alreadysupplementedwith 1%b-mercaptoethanol) to each sample and pipet up and down several times to mix thoro

14 5. Add700µloflowstringencywashsolutiontoeachwelloftheRNAbinding plate. Centrifuge for 2 min at 1,500 x g. Discard the low stringen

15 Section 9 Troubleshooting Guide Problem Possible Cause Recommended Solution Difficulty achieving Purge step in protocol If sealing of well

16 Problem Possible Cause Recommended Solution Low or highly variable Elution solution applied Apply elution solution eluate volumes among to

17 Problem Possible Cause Recommended Solution Clogging of RNA Excessive amount of Reduce volume of binding plate starting material per well

18 Problem Possible Cause Recommended Solution Total RNA prep Incorrect use of wash Add the appropriate performs poorly in  solutions volum

19 Section 10 OrderingInformation Catalog # Description 732-6800 Aurum™ Total RNA 96 Kit 732-6470 Aurum™ Vacuum Manifold 732-6820 Aurum™

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Table of Contents Section 1 Introduction...1 Section 2 Kit Components ...1 Section 3 Storage

Section 1 Introduction The Aurum™ Total RNA 96 kit rapidly purifies up to 192 total RNA samples from biological samples (e.g. mammalian cells, yea

Section 3 Storage Conditions All kit components (including lyophilized DNase I) should be stored at room temperature. Store reconstituted DNase I

Section 5 BeforeUsingthe Aurum™ Total RNA 96 Kit Please read the following guidelines before proceeding with the total RNA purification. Start

Table 1. Yield (per well) of total RNA from various samples using the Aurum™ Total RNA 96 kit. Starting Material Avg. Yield (µg)* Cultured cell

• Vendors of lyticase, which is used to partially degrade the cell walls of yeast cells, may have different definitions of the enzyme’s activity.