Cuisinart SG-10A Troubleshooting Guide

QIAprep®Miniprep HandbookFor purification of molecular biology grade DNA PlasmidLarge plasmids (>10 kb)Low-copy plasmids and cosmidsPlasmid DNA pre

10 QIAprep Miniprep Handbook 12/2006IntroductionThe QIAprep Miniprep system provides a fast, simple, and cost-effective plasmidminiprep method for r

QIAprep Miniprep Handbook 12/2006 11Applications using QIAprep purified DNA Plasmid DNA prepared using the QIAprep system is suitable for a variety

12 QIAprep Miniprep Handbook 12/2006LyseBlue is a color indicator which provides visual identification of optimum buffer mix-ing. This prevents comm

QIAprep Miniprep Handbook 12/2006 13QIAprep SpinFigure 1 10 µg pUC18 DNA was purified using the QIAprep Spin protocol and eluted with the indicated

Table 1. Effect of Different Compositions of Growth Medium LB on DNA YieldCulture media YieldLB (containing 10 g/liter NaCl) 11.5 µgLB (containing 5 g

QIAprep Miniprep Handbook 12/2006 15Important Notes Please read the following notes before starting any of the QIAprep procedures.Growth of bacteria

16 QIAprep Miniprep Handbook 12/2006Buffer notes Add the provided RNase A solution to Buffer P1, mix, and store at 2–8°C. Add ethanol (96–100%) to

QIAprep Miniprep Handbook 12/2006 17Table 2. Regulation of Vacuum Pressures for QIAprep Multiwell ProceduresProcedure Vacuum Module used for Vacuum

18 QIAprep Miniprep Handbook 12/2006Multichannel pipet recommendationsMany steps of the QIAprep 8 procedure and the QIAprep 8 and 96 Turbo procedure

To ensure consistent performance, do not apply silicone or vacuum grease to anypart of a QIAvac manifold. The spring lock on the top plate and the s

Trademarks: QIAGEN®, QIAprep®, BioRobot®, R.E.A.L.®, TurboFilter®(QIAGEN); DH5α™ (Invitrogen); pBluescript®(Stratagene); pGEM®(Promega Corp.);Beckman®

20 QIAprep Miniprep Handbook 12/2006QIAvac 6S Manifold108957124636Figure 4 Components of the QIAvac 6S manifold.1. QIAvac base, which holds a waste

QIAprep Miniprep Handbook 12/2006 21QIAprep Spin Procedurein microcentrifuges on vacuum manifoldsResuspendLyseNeutralizePelleted bacteriaPure plasmi

QIAprep Spin22 QIAprep Miniprep Handbook 12/2006Protocol: Plasmid DNA Purification Using the QIAprepSpin Miniprep Kit and a Microcentrifuge This pro

QIAprep SpinQIAprep Miniprep Handbook 12/2006 235. Apply the supernatants from step 4 to the QIAprep spin column by decanting or pipetting.6. Centri

QIAprep Spin24 QIAprep Miniprep Handbook 12/2006Protocol: Plasmid DNA Purification Using the QIAprepSpin Miniprep Kit and a Vacuum ManifoldThis prot

QIAprep SpinQIAprep Miniprep Handbook 12/2006 25During centrifugation, prepare the vacuum manifold and QIAprep spin columns:QIAvac 24 Plus (see page

9. Transfer the QIAprep spin columns to a microcentrifuge tube. Centrifuge for 1 min.Important: This extra spin is necessary to remove residual Buffer

QIAprep Miniprep Handbook 12/2006 27QIAprep 8 ProcedurePelleted bacteriaResuspendLyseNeutralizeBindPure plasmid DNAEluteWashVacuumVacuumVacuumQIApre

QIAprep 828 QIAprep Miniprep Handbook 12/2006Protocol: Plasmid DNA Purification Using the QIAprep8 Miniprep Kit This protocol is designed for purifi

QIAprep 8QIAprep Miniprep Handbook 12/2006 29Make sure the QIAvac 6S is assembled correctly before loading. Load the super-natants promptly onto the

QIAprep Miniprep Handbook 12/2006 3ContentsKit Contents 4Storage 7Quality Control 7Product Use Limitations 7Product Warranty and Satisfaction Guaran

QIAprep 8 Turbo30 QIAprep Miniprep Handbook 12/2006QIAprep 8 and 96 TurboProcedureResuspendLyseNeutralizePure plasmid DNABindEluteWashPelleted bacte

QIAprep 8 TurboQIAprep Miniprep Handbook 12/2006 31Protocol: Plasmid DNA Purification Using the QIAprep8 Turbo Miniprep Kit This protocol is designe

QIAprep 8 Turbo32 QIAprep Miniprep Handbook 12/20065. Switch off vacuum, and ventilate the QIAvac 6S slowly. Discard the TurboFilterstrips. Transfer

QIAprep 96 TurboQIAprep Miniprep Handbook 12/2006 33Protocol: Plasmid DNA Purification Using the QIAprep96 Turbo Miniprep Kit This protocol is desig

QIAprep 96 Turbo34 QIAprep Miniprep Handbook 12/20064. Remove the tape from the block. Pipet the lysates from step 3 (850 µl per well) intothe well

QIAprep 96 TurboQIAprep Miniprep Handbook 12/2006 3510a. For elution into provided collection microtubes: Replace waste tray with the blue collectio

36 QIAprep Miniprep Handbook 12/2006Troubleshooting GuideThis troubleshooting guide may be helpful in solving any problems that may arise. The scien

QIAprep Miniprep Handbook 12/2006 37Comments and suggestionsb) RNase A digestion Ensure that RNase A is added to Buffer P1 before use.omittedc) RNa

38 QIAprep Miniprep Handbook 12/2006Comments and suggestionsRNA in the eluatea) RNase A digestion Ensure that RNase A is added to Buffer P1 before

QIAprep Miniprep Handbook 12/2006 39Appendix A: Background InformationGrowth of bacterial culturesPlasmids are generally prepared from bacterial cul

Kit ContentsQIAprep Spin Miniprep Kit (50) (250)Catalog no. 27104 27106QIAprep Spin Columns 50 250Buffer P1 20 ml 73 mlBuffer P2 20 ml 73 mlBuffer N3

40 QIAprep Miniprep Handbook 12/2006Host strainsMost E. colistrains can be used successfully to isolate plasmid DNA, although the strainused to prop

QIAprep Miniprep Handbook 12/2006 41Table 6. Concentrations of Commonly Used AntibioticsStock solutions Working concentrationAntibiotic Concentratio

42 QIAprep Miniprep Handbook 12/2006Preparation of cell lysatesBacteria are lysed under alkaline conditions. After harvesting and resuspension, the

QIAprep Miniprep Handbook 12/2006 43Appendix B: Agarose Gel Analysis of Plasmid DNAThe QIAprep Miniprep procedure can be analyzed using agarose gel

44 QIAprep Miniprep Handbook 12/2006Appendix C: Special Applications Purification of low-copy plasmids and cosmidsAll QIAprep miniprep protocols in

QIAprep Miniprep Handbook 12/2006 45References1. Vogelstein, B., and Gillespie, D. (1979) Preparative and analytical purification of DNA from agaro

46 QIAprep Miniprep Handbook 12/2006Ordering InformationProduct Contents Cat. no.QIAprep Spin For 50 plasmid minipreps: 27104Miniprep Kit (50) 50

QIAprep Miniprep Handbook 12/2006 47Ordering InformationProduct Contents Cat. no.QIAprep 96 Turbo For 4 x 96 plasmid minipreps, 962141BioRobot Kit

48 QIAprep Miniprep Handbook 12/2006Ordering InformationProduct Contents Cat. no.QIAvac 6S Vacuum manifold for processing 1–6 19503QIAGEN 8-well st

QIAprep Miniprep Handbook 12/2006 49Ordering InformationProduct Contents Cat. no.Individual Buffers and accessoriesBuffer N3 500 ml Buffer N3 19064B

QIAprep Miniprep Handbook 12/2006 5QIAprep 8 Turbo Miniprep Kit (10) (50)Catalog no. 27152 27154TurboFilter®8 Strips 10 50QIAprep 8 Strips 10 50Buf

50 QIAprep Miniprep Handbook 12/2006Notes

QIAprep Miniprep Handbook 12/2006 51QIAGEN Distributors and ImportersPlease see the back cover for contact information for your local QIAGEN office.

WWW.QIAGEN.COM1043788 12/2006Australia Orders 03-9840-9800 Fax 03-9840-9888 Technical 1-800-243-066Austria Orders: 0800/28-10-10 Fax: 0800/28-10

Please tear off hereBench Protocol: QIAprep Spin Miniprep Kit Using a Vacuum ManifoldThis protocol is designed for the purification of up to 20 µg hig

Bench Protocol: QIAprep Spin Miniprep Kit Using a MicrocentrifugeThis protocol is designed for the purification of up to 20 µg high-copy plasmid DNAfr

6 QIAprep Miniprep Handbook 12/2006QIAprep 96 Turbo Miniprep Kit (4) (24)Catalog no. 27191 27193TurboFilter 96 Plates 4 24QIAprep 96 Plates 4 24Buf

QIAprep Miniprep Handbook 12/2006 7StorageQIAprep Miniprep Kits should be stored dry at room temperature (15–25°C). Kits canbe stored for up to 12 m

8 QIAprep Miniprep Handbook 12/2006Technical AssistanceAt QIAGEN we pride ourselves on the quality and availability of our technical support.Our Tec

QIAprep Miniprep Handbook 12/2006 9Safety InformationWhen working with chemicals, always wear a suitable lab coat, disposable gloves,and protective