mn guide to plasmid purification - 3 jjnucleobond ® xtra midi • nucleobond xtra maxi...
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www.mn-net.comMACHEREY-NAGEL GmbH & Co. KG Neumann-Neander-Str. 68 52355 Dren GermanyFrance:MACHEREY-NAGEL EURLTel.: +33 388 68 22 68Fax: +33 388 51 76 88E-mail: sales-fr@mn-net.com
Switzerland:MACHEREY-NAGEL AGTel.: +41 62 388 55 00Fax: +41 62 388 55 05E-mail: sales-ch@mn-net.com
Germanyand international:Tel.: +49 24 21 969-0Fax: +49 24 21 969-199E-mail: info@mn-net.com
MACHEREY-NAGEL
EN ISO 9001: 2008CERTIFIED
USA:MACHEREY-NAGEL Inc.Tel.: +1 484 821 0984Fax: +1 484 821 1272E-mail: sales-us@mn-net.com
Plas
mid
Pur
ifica
tion
Gui
de Plasmid purification productsfrom MACHEREY-NAGELMN guide to plasmid purification Find the optimal solutionNucleoBond
NucleoSpin
Superior yieldsOutstanding puritiesTime-saving procedures for reliable downstream applications
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NucleoSpinSilica-membrane technology .......................................NucleoSpin kits see page 6
DNAcontaminants
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 NucleoBondFinalizer
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) OptimalcolumndesignandinsertedfiltersavoidinconvenientsyringesXX Highest yield LyseControl for visualization of completed alkaline lysis Typically250gforMidiand1000gforMaxiXX High purity transfection-grade plasmid DNA
Established anion-exchange technologyXX NucleoBond Xtra Plus kits for super high-speed version
NucleoBondFinalizerforomittingtime-consumingcentrifugationafterDNAprecipitation
Product at a glanceSample material Midi