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de Plasmid purification productsfrom MACHEREY-NAGEL
MN guide to plasmid purification Find the optimal solutionNucleoBond®
NucleoSpin®
Superior yieldsOutstanding puritiesTime-saving procedures… for reliable downstream applications
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NucleoSpin®
Silica-membrane technology .......................................NucleoSpin® kits see page 6DNAcontaminants
Sample lysis, release of DNA from cells, tissue
DNA is bound to the silica membrane under high-salt conditions Interaction between DNA (hydrate shell is reversibly removed by chaotropic salt) and silica membrane
Contaminants are washed away under high-salt and / or ethanolic conditions to keep the DNA bound to themembrane
DNA is eluted in low-salt buffer or water, DNA is ready to use for downstream applications
Principle of binding: Removing of hydrate shell Hydrogen bonds / Salt bridges
Principle of elution: Reconstitution of hydrate shell
Features / Results •SequencingandPCR-gradeplasmidDNA •Noalcoholprecipitationnecessary •Fastandeasyprocedure
MN technologies for plasmid purificationNucleoBond® Anion-exchange technology ........................................NucleoBond® kits see page 3
DNAcontaminants
DNA is bound to the anion-exchanger matrix under low-pH conditions Interaction between positively charged anion-exchanger group and negatively charged DNA backbone
Stringent washing with increasing salt concentration to remove contaminants
DNA is eluted with high-pH buffer
Desalting / Concentration: Alcohol precipitation of eluted DNA DNA is collected by centrifugation or by using the NucleoBond®Finalizer
Si spacer NH
CH3
OHO
CH2
O
P
O
O
O
anion-exchangergroup MAE
DNA backbone
binding
Principle of binding: Ionic bond
2
3
O
CH
O
P
O
O
O
elution of DNA
anion-exchangergroup MAE
DNA backbone
pH shift
Si spacer N
CH
OH
Principle of elution: pH shift
Features / Results •Ultra-pure,transfection-gradeplasmidDNA •Thenewgenerationofanionexchangers •Xtrafast,Xtrahighyield,Xtraconvenient
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JJNucleoBond® Xtra Midi • NucleoBond® Xtra Maxi Superior plasmid Midi and Maxi kits – transfection-gradeXX Highest speed – 30 min (Midi) / 35 min (Maxi) Optimalcolumndesignandinsertedfilters–avoidinconvenientsyringesXX Highest yield – LyseControl for visualization of completed alkaline lysis Typically250μgforMidiand1000μgforMaxiXX High purity – transfection-grade plasmid DNA
Established anion-exchange technologyXX NucleoBond ® Xtra Plus kits for super high-speed version
* Yield of plasmid DNA is slightly lower due to residual DNA remaining on the desalting tool (compared to kits without desalting tool).
Highest yield in less time – comparison to anion-exchange competitor kits
PlasmidDNAwas isolated followingeachmanufacturer’sprotocolusingmaximumculturevolume.PlasmidDNAyield () was determined after precipitation.ThecomparisonshowstheresultsofthekitswithNucleoBond®Finalizerasdesaltingtool(Fig.A)andwithoutdesaltingtool(Fig.B).
Customer testimonial„We have tested many of the market leaders in anion-exchange purification, and the MACHEREY-NAGEL NucleoBond Xtra® kits excel in ease of use, speed of preparation, yield, and DNA quality.“EnriqueSaez,PhD,TheGenomicsInstituteofNovartisResearchFoundation,SanDiego,CA,USA
NucleoBond ® Xtra Maxi Plus (incl. Finalizer) 10/50 740416.10/.50
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JJNucleoBond® Xtra Midi EF • NucleoBond® Xtra Maxi EF Superior plasmid kit for transfection of sensitive cell lines – endotoxin-freeXX Highest purity – endotoxin-free transfection of sensitive cell lines
Patented endotoxin removalXX Highest speed – 45 min (Midi) / 50 min (Maxi)
No extra incubation for endotoxin removal Optimalcolumndesignandinsertedfilters–avoidinconvenientsyringesXX Highest yield – typically 250 μg for Midi and 1000 μg for Maxi Improvedsilicamaterial–patenteddesign!XX NucleoBond ® Xtra Plus EF kits for super high speed version
Customer testimonial„Our lab is constantly doing transfections and we use no other kit but the MACHEREY-NAGEL NucleoBond Xtra® Maxi EF. It is easy and fast to use and always gives us great, consistent results so that we spend less time prepping plasmids and more time on experiments that matter.“ MollyRomine,Pediatrics,WashingtonUniversitySchoolofMedicine,St.Louis,MO,USA
Ordering informationProduct Preps REF
NucleoBond ® Xtra Midi EF 10/50 740420.10/.50
NucleoBond ® Xtra Midi Plus EF (incl. Finalizer) 10/50 740422.10/.50
NucleoBond ® Xtra Maxi EF 10/50 740424.10/.50
NucleoBond ® Xtra Maxi Plus EF (incl. Finalizer) 10/50 740426.10/.50
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JJNucleoBond® Xtra BAC Kit for large construct DNA XX High yield – up to 150 μg BAC DNA
Optimal silica material for large constructs LyseControlforvisualizationofefficientalkalinelysisXX High speed – 75 minutes per prep OptimalcolumndesignandinsertedfiltersforhighflowratesandparallellysateclearingandloadingXX High purity – transfection-grade BAC DNA
Established anion-exchange technology
Product at a glanceSample material 250–750mLE. coli cultureVectorsize <300kbpTypicalyield 10–150μgPreparation time 75min/prepApplications Transfection,cloning,restrictionanalyses,sequencing
Application datSuperior yield in less time – comparison to competitor kits
BACDNA (300kbp)was isolated in triplicate from500mLE. coliDH5αusingNucleoBond®XtraBACandcompetitorproducts(QandI).Afterprecipitation,BACDNAwasreconstitutedin1000μLTEbufferand5μLofeachsamplewasusedforanalysisona1%TAE-agarosegel.Obtained yields:MN:150μg,Q:44μg,I:75μg
Ordering informationProduct Preps REF
NucleoBond ® Xtra BAC 10/25 740436.10/.25
JJNucleoBond® 96 Xtra EF High-throughput isolation of endotoxin-free plasmid DNAXX Highest purity – endotoxin-free transfection of sensitive cell lines
Patented endotoxin removalXX NucleoBond® Filter Plate for convenient filtration of bacterial lysates
Processing under vacuum or by centrifugation InnovativeMNWashPlateminimizesriskofcross-contaminationXX NucleoBond® Finalizer Plate to speed up DNA precipitation
Product at a glanceSample material 1–5mLE. coli cultureVectorsize <15kbp
10–50μg(5mLLB/TBinglasstubes)Preparation time 120min/plateApplications Transfection,cloning,plasmidbanks,etc.
Ordering informationProduct Preps REF
NucleoBond ® 96 Xtra EF 1x96/4x96 740430.1/.4
MN Q I
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JJNucleoSpin® Plasmid · NucleoSpin® Plasmid (NoLid) High-yield plasmid Mini prep – molecular biology- / sequencing-grade XX Highest yield Upto50μgplasmidDNAXX Highest quality Phredscoreof20(upto1000bp)XX Fast procedure 6Miniprepsin25minXX Choose between columns with or without a lid
Product at a glanceSample material 1–5mLE. coli cultures (standard protocol)
JJNucleoSpin® Plasmid EasyPure High-speed plasmid Mini prep – molecular biology- / sequencing-gradeXX Highest speed – one combined washing and drying step 6Miniprepsin14minXX LyseControl for visualization of completed alkaline lysis Obtainupto35μgplasmidDNAXX Now with Liquid RNase
Easy handling without dissolving
Product at a glanceSample material 2–10mLE. coli cultures Vectorsize <15kbpTypicalyield 15–30μgElution volume 50μLBinding capacity 35μg
NucleoSpin®8FilterStrips/96FilterPlate OptimizedfiltermaterialminimizestheriskoffiltercloggingXX Minimized risk of cross-contamination MNWashPlateXX Processing under vacuum or by centrifugation*
Suitable for manual and automated processingXX NucleoSpin® 8 / 96 Plasmid Core Kit: Kitswithbasiccontentfocusedonautomationplatforms–additionalaccessoriescanbecombinedasneeded
Product at a glanceSample material 1–5mLE. coli cultureVectorsize <15kbpTypicalyield 4–6μg/mLE. coli cultureElution volume 75–150μLPreparation time 45min / 6stripsor1plateBinding capacity 20μgApplications Sequencing,cloning,PCR,restrictionanalysis,transformation,enzymaticmodifications