Sample Data Sheet
Contents
1. User s Manual Product Serum DNA Isolation Kit Catalog Number K5018100 Shipping Condition Room temperature Introduction The Serum DNA Isolation Kit is designed for the purification of genomic DNA fragment from serum plasma and bio fluid in a spin column format No phenol chloroform extraction no Protease and no precipitation steps are involved The sample addition and washing steps can be performed using compatible vacuum manifold while the final elution of the DNA product are performed using a table top centrifuge The samples are first treated in the binding buffer that contains the denaturant guanidine HCI and Triton X100 Ethanol is then added to the samples which are then added to the filter spin columns This step facilitates the binding of DNA to the filter matrix Under these conditions the DNA binds to the membrane while other contaminants are washed through The columns are then washed to further remove protein buffer components and other contaminants using two ethanol containing wash buffers and the final genomic DNA product is eluted in TE The final DNA product can be used directly for quantitative PCR and other downstream applications Feature e No phenol chloroform e No protease e No precipitation e Total lt 15 min e Sample range 200 ul Kit Contents 1 Lysis Binding Buffer K5018100 1 21ml RT 2 Wash Buffer 1 K5018100 2 26mi RT RT 4 TE oo K5od8ioo 4 fiom RT K5018100 5 100units RT Storage2. Conditions All of contents of the Serum DNA Isolation Kit including the buffers should be stored at room temperature The kit is stable for one year under these conditions RT l RT RT RT RT Technical Assistance Please refer any technical questions to TechSupport biochain com Important Notes Before Using The Serum DNA Isolation Klit Sample Size and Type The Serum DNA Isolation Kit can be used to isolate genomic DNA using a spin column format DNA can be isolated quantitatively from serum plasma or other bio fluid Buffer Concentrates Wash buffers 1 and 2 are provided as concentrates that require the addition of 100 ethanol to them before use Reagents and Equipment to be Supplied by the User e Pipetteman multichannel pipettors desirable e 1 5 ml tubes UK amp Rest of World Switzerland Deutschland United States Centro Nord Sud 2E CH 6934 Bioggio Lugano Tel 41 0 91 604 55 22 Fax 41 0 91 605 17 85 Bockenheimer Landstr 17 19 60325 Frankfurt Main Tel 49 0 69 779099 Fax 49 0 69 13376880 23591 El Toro Rd Suite 167 Lake Forest CA 92630 Tel 1 800 987 0985 Fax 1 949 265 7703 184 Milton Park Abingdon OX14 4SE Oxon UK Tel 44 0 1235 828 200 Fax 44 0 1235 820 482 amsbio e Disposable gloves e 100 ethanol e Distilled deionized water e A table top centrifuge capable of providing gt 13k rpm rotor Protocol Before starting The wa
3. d are therefore difficult to determine with a spectrophotometer or other DNA detection method Quantitative amplification methods such as PCR are recommended for determination of yield Related Products EZ Blood DNA 96 Kit Cat Z7040008 EZ DNA 96 Kit Cat Z7040006 Genomic DNA Extraction Kit Cat K5016005 Blood DNA Isolation Kit Cat K5017100 Trouble Shooting Problem Little or no DNA eluted Remove all traces of supernatant before beginning All buffers must be at room temperature Ensure that vacuum draws all liquid through filter membrane at each step Measure final elution volume ensure adequate final elution from final centrifugation steps Filters clog Too much DNA cells used Reduce sample size Filters tear Reduce centrifugation speed UK amp Rest of World Switzerland Deutschland United States Centro Nord Sud 2E CH 6934 Bioggio Lugano Tel 41 0 91 604 55 22 Fax 41 0 91 605 17 85 Bockenheimer Landstr 17 19 60325 Frankfurt Main Tel 49 0 69 779099 Fax 49 0 69 13376880 184 Milton Park Abingdon OX14 4SE Oxon UK Tel 44 0 1235 828 200 Fax 44 0 1235 820 482 amsbio 23591 El Toro Rd Suite 167 TOF ET Lake Forest CA 92630 into amsbio c Tel 1 800 987 0985 Fax 1 949 265 7703 Jill Serum DNA Isolation Kit Experienced Users Miniprotocol 200 ul serum amp 200 ul binding buffer mix 5 min RT 500 ul wash buffer 1 spi
4. n 13k rpm 1 min 600 ul wash buffer 2 spin 13k rpm 1 min spin 13k rom 1 min transfer the column onto collection tube 50 ul of TE incubate 1 min spin 13k rpm 1 min a ee FOR RESEARCH USE ONLY UK amp Rest of World Switzerland Deutschland United States amsbio 184 Milton Park Abingdon OX14 4SE Oxon UK Tel 44 0 1235 828 200 Fax 44 0 1235 820 482 Centro Nord Sud 2E CH 6934 Bioggio Lugano Tel 41 0 91 604 55 22 Fax 41 0 91 605 17 85 Bockenheimer Landstr 17 19 60325 Frankfurt Main Tel 49 0 69 779099 Fax 49 0 69 13376880 23591 El Toro Rd Suite 167 Lake Forest CA 92630 Tel 1 800 987 0985 Fax 1 949 265 7703
5. sh buffer 1 concentrate requires the addition of 26 ml of 100 ethanol before it can be used while the wash buffer 2 concentrate requires that 48 ml of 100 ethanol is added to it before use Both of the wash buffers are stable for one year after the addition of ethanol 1 Transfer up to 200 ul of serum or plasma into a 1 5 ml tube Add 200 ul of binding buffer per tube Pipette up and down until mix well cap the tube and incubate at room temperature for 10 min 2 Add the contents to the filter column Spin down the column at 13k rom for one minute and discard the spin through liquid 3 Wash the column by adding 500 ul wash buffer 1 which contains the added ethanol per column and spin the column as above condition 4 Wash the column once by adding 600 ul wash buffer 2 which contains the added ethanol and spin the column as above 5 Discard the liquid in the collection tube and spin the column one more minutes as above condition 6 Place the column onto a new 1 5 ml tube for DNA sample elution Add 50 ul of TE per column and wait one min and then centrifuge at 13k rom for 1 min to elute the final DNA product The use of 30 ul elution rather than 50 ul elution steps will provide you with a more concentrated DNA product however the absolute yield of DNA will be reduced and the intrawell variation increased Kit Performance Yield of DNA fragment isolated from biological samples using the EZ Serum DNA Kit is normally below 1 ug an
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