Bio-rad PureZOL™ RNA Isolation Reagent Instrukcja Użytkownika

PureZOL™RNA IsolationReagentInstruction ManualCatalog #732-6890For technical support,call your local Bio-Rad office, or in the US, call 1-800-4BIORAD

recommended to disrupt the cell walls of yeast and bacteria.Bacteria and yeast lysate can also be heated to 55°C for 10minutes prior to adding chlorof

100 mg of tissue and transfer the sample into a suitable sizedtube for disruption and homogenization. Add 1 ml of PureZOLreagent to every 50–100 mg of

top layer. Carryover of the solid debris can cause column clog-ging and affect RNA sample purity.3. Add 0.2 ml of chloroform per 1 ml of PureZOL used

8. Centrifuge at 12,000 x g for 10 minutes at 4°C.9. The RNA will appear as a white pellet on the side and bottom of the tube. Carefully discard the s

Section 6TroubleshootingProblem Possible Cause Recommended SolutionIncomplete Lysate was not mixed Once chloroform is added, separation of properly a

Problem Possible Cause Recommended SolutionLow yield the pellet. If necessary, (continued) incubate the resuspendedpellet for 10 minutes at55–60°C and

Problem Possible Cause Recommended SolutionLow yield Insufficient amount of Scale up the amount of(continued) PureZOL used for PureZOL used according

Problem Possible Cause Recommended SolutionRNA is Cells grown in either For cells grown in monolayer,degraded monolayer or aspirate the growth mediu

Problem Possible Cause Recommended SolutionGenomic DNA Phase separation not Make sure that centrifugationcontamination performed at the right step i

Section 8Ordering InformationCatalog # Description732-6890 PureZOL RNA isolation reagent, 100 mlRelated ProductsAurum Total RNA Kits for Isolation of

Table of ContentsSection 1 Introduction...1Kit Components...

Legal NoticesPureZOL RNA isolation reagent is subject to US Patent 5,346,994.Speed Vac is a trademark of Sevant Instruments, Inc.1810001481A.qxp 3/7/

Bio-Rad Laboratories, Inc.2000 Alfred Nobel Dr.Hercules, CA 94547 USA(510) 741-10001-800-424-672310001481 Rev A10001481A.qxp 3/7/2005 7:05 AM Pag

Section 1IntroductionPureZOL RNA isolation reagent is intended for the extraction oftotal RNA from animal and plant tissues, cultured mammaliancells,

Kit ComponentsCatalog # 732-6890 – PureZOL RNA isolation reagent, 100 ml Storage and StabilityPureZOL RNA isolation reagent is shipped at room tempera

Section 2Materials and Equipment Required(Not Provided)Microcentrifuges (capable of >12,000 x g) — at 4°C and at roomtemperature95–100% ethanol, AC

Section 3Maintaining an RNase-Free EnvironmentTo avoid introducing RNases, great care must be taken in handling the reagents and purified RNA samples.

Solutions that are prepared by the user should be treated withDEPC to inactivate RNases. See instructions below on how to prepare DEPC-treated solutio

Section 4Recommendations for Best Results• When isolating RNA from small sample sizes (<500,000 cells or<10 mg of tissue), lyse or homogenize in

Section 5ProtocolCarry out all steps at room temperature unless otherwise indicated.RNase-free disposable polypropylene tubes should be usedthroughout