Bio-Spin-6-column.pdf

1、nucleic acid purificationBio-Spin6 and 30 ColumnsAutoclavability Bio-Spin columns,Bio-Gel P gel,and collection tubes arecompletely autoclavable at 121C for 30 min at pH 6.08.0.Chemical StabilitypH 210,common aqueous buffers,formamide,dilute organicacids,alcohol,20%(v/v)other chaotropic agents,deterg

2、ents.StorageStore at 4C.Do not freeze.Instructions for Use1Invert the column sharply several times to resuspend thesettled gel and remove any bubbles.2Snap off the tip and place column in a 2.0 ml microcentrifugetube(included).Remove cap.Allow the excess packing bufferto drain by gravity to top of g

3、el bed.(If column does not beginto flow,push cap back into column and remove).Discard thedrained buffer,then place the column back into 2 ml tube.However,if placing column into 12 x 75 mm test tube,centrifuge immediately.3Centrifuge for 2 min in a swinging bucket centrifuge at 1,000 x g(see Centrifu

4、gation Notes section)to remove the packingbuffer.Discard the buffer.4Place the column in a clean 2.0 ml microcentrifuge tube or 12 x 75 mm test tube.Carefully apply the sample(20100 l)directly to the center of the column.Application of more or lessthan the recommended sample volume may decrease colu

5、mnperformance.5After loading sample,centrifuge the column for 4 min at 1,000 x g.6Following centrifugation,the purified sample is now in Tris orSSC buffer.Molecules smaller than the columns exclusion limitwill be retained.7Properly dispose of the used column.Buffer ExchangeThe gel in the Bio-Spin co

6、lumns is suspended in either SSCbuffer,pH 7.0,or Tris buffer,pH 7.4.The gel matrix iscompatible with most aqueous buffers.Buffer exchange canbe achieved using the following procedure.IntroductionBio-Spin chromatography columns are ready to use for rapidand efficient cleanup and purification of nucle

7、ic acids andproteins using a swinging bucket centrifuge.Bio-Spin 6 and 30 Columns Remove dye terminators Remove unincorporated nucleotides Desalting and buffer exchangeThe columns are packed with special grades of Bio-GelPpolyacrylamide P-6 or P-30 gel matrices manufacturedspecifically for Bio-Rad s

8、pin columns.This unique gelproduces very efficient,noninteractive size separations.Werecommend Bio-Spin 6 Tris columns for buffer exchange anddesalting applications,while Bio-Spin 30 Tris columns areoptimal for removal of unincorporated nucleotides.Bio-Spincolumns are suitable for use with 2.0 ml mi

9、crocentrifuge tubesor 12 x 75 test tubes and are completely autoclavable.Technical InformationGel MatrixBio-Gel P-6 or P-30 polyacrylamide gel suspended in 1.0 ml ofbuffer.BuffersSSC buffer (150 mM sodium chloride,17.5 mM sodium citrate,pH 7.0)with 0.02%sodium azide.Tris buffer(10 mM Tris-HCl,pH 7.4

10、)with 0.02%sodium azide.Sample Application VolumesNucleic acids,proteins,and peptides,20100 l.Exclusion Limits Bio-Gel P-6 gel:5 base pairs(nucleic acids)or molecularweight 6,000(proteins,peptides).Bio-Gel P-30 gel:20 base pairs(nucleic acids)or molecularweight 40,000(proteins,peptides).Expected Ret

11、ention and Recovery Up to 99%retention of unincorporated nucleotides.Up to 95%recovery of applied DNA.Centrifuge Type Swinging bucket centrifuge with a centrifugal force of 1,000 x g.Life ScienceGroupWeb Site www.bio- U.S.(800)4BIORAD Australia 02 9914 2800 Austria(01)-877 89 01 Belgium 09-385 55 11

12、 Canada(905)712-2771 China 86-10-62051850/51 Denmark 45 39 17 99 47 Finland 358(0)9 804 2200 France 01 43 90 46 90 Germany 089 318 84-0 Hong Kong 852-2789-3300 India(91-11)461-0103 Israel 03 951 4127 Italy 39-02-216091 Japan 03-5811-6270 Korea 82-2-3473-4460 Latin America 305-894-5950 Mexico 514-221

13、0 The Netherlands 0318-540666 New Zealand 64-9-4152280 Norway 22-74-18-70 Russia 7 095 979 98 00 Singapore 65-2729877 Spain 34-91-661-7085 Sweden 46(0)8-55 51 27 00 Switzerland 01-809 55 55 United Kingdom 0800-181134 LIT-507 Rev GBio-RadLaboratories1Follow steps 1,2,and 3 in the Instructions for Use

14、 section.2Apply the new buffer in 500 l aliquots.After each applicationof new buffer,let the buffer drain out by gravity,or centrifugethe column for 1 min to remove the buffer.Discard buffer fromcollection tube.Repeat as required.Three washes result in99%of the buffer being exchanged.Four washes res

15、ult in99.9%of buffer exchanged.3Sample can now be applied to the column as directed in steps4 through 7 in the Instructions for Use section.Centrifugation NotesBio-Spin columns fit 2.0 ml microcentrifuge tubes or 12 x 75 mmtest tubes for sample collection during centrifugation.Use the2.0 ml microtub

16、es provided with the columns for the initialcolumn equilibration step.Swinging bucket centrifuges capable of generating a minimumforce of 1,000 x g are suitable for Bio-Spin column use.Thegravitational force created at a particular revolution speed is afunction of the radius of the microcentrifuge r

17、otor.Consult theswinging bucket centrifuge instruction manual for conversioninformation from revolutions per minute(RPM)to centrifugalor g-force.Alternatively,to calculate the speed in RPMrequired to reach a gravitational force of 1,000 x g,use thefollowing equation:RCF(x g)=(1.12 x 10-5)x(RPM)x 2 x

18、 r where RCF is the relative centrifugal force,r is the radius incentimeters measured from the center of the rotor to themiddle of the Bio-Spin column,and RPM is the speed ofthe rotor.SterilizationIf a sterile Bio-Spin column is required,autoclave the columnat 121C for 2030 min.If exchanging buffers

19、,the buffer pHin the column should be in the range of 6.0 to 8.0 priorto autoclaving.Ordering InformationCatalog#Product DescriptionBio-Spin Columns with Bio-Gel P-6 in Tris Buffer732-6227Bio-Spin 6 Tris Columns,25732-6228Bio-Spin 6 Tris Columns,100732-6229Bio-Spin 6 Tris Columns,1,000Bio-Spin Colum

20、ns with Bio-Gel P-30 in Tris Buffer732-6231Bio-Spin 30 Tris Columns,25732-6232Bio-Spin 30 Tris Columns,100732-6233Bio-Spin 30 Tris Columns,1,000Bio-Spin Columns with Bio-Gel P-6 in SSC Buffer732-6002Bio-Spin 6 SSC Columns,25Bio-Spin Columns with Bio-Gel P-30 in SSC Buffer732-6006Bio-Spin 30 SSC Colu

21、mns,25Micro Bio-Spin Columns with Bio-Gel P-6 in Tris Buffer732-6221Micro Bio-Spin 6 Tris Columns,25732-6222Micro Bio-Spin 6 Tris Columns,100732-6225Micro Bio-Spin 6 Tris Columns,1,000Micro Bio-Spin Columns with Bio-Gel P-30 in Tris Buffer732-6223Micro Bio-Spin 30 Tris Columns,25732-6224Micro Bio-Sp

22、in 30 Tris Columns,100732-6226Micro Bio-Spin 30 Tris Columns,1,000Micro Bio-Spin Columns with Bio-Gel P-6 in SSC Buffer732-6200Micro Bio-Spin 6 SSC Columns,25732-6201Micro Bio-Spin 6 SSC Columns,100Micro Bio-Spin Columns with Bio-Gel P-30 in SSC Buffer732-6202Micro Bio-Spin 30 SSC Columns,25732-6203

23、Micro Bio-Spin 30 SSC Columns,100RNase-Free Micro Bio-Spin Columns with Bio-Gel P-30 in Tris Buffer732-6250Micro Bio-Spin 30 Tris Columns,RNase free,25732-6251Micro Bio-Spin 30 Tris Columns,RNase free,100Empty Columns732-6008Empty Bio-Spin Columns,100732-6204Empty Micro Bio-Spin Columns,100PCR React

24、ion Mixture Purification732-6300PCR Kleen Spin Columns,25732-6301PCR Kleen Spin Columns,100DNA Recovery from Agarose Gels732-6165Freeze-N-Squeeze Spin Columns,25732-6166Freeze-N-Squeeze Spin Columns,100732-6160Quantum Prep Gel Slice KitSEQueaky Kleen Terminator Removal Kit732-6260SEQueaky Kleen,2x96732-6261SEQueaky Kleen,10 x96