AllPrep DNA/RNA Micro プロトコールとトラブルシューティング ( /2008)

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1 July 2007 AllPrep DNA/RNA Micro DNA RNA 5 x mg Sample & Assay Technologies

2 DNA RNA 3 DNA RNA 11 DNA RNA AllPrep DNA/RNA Micro 07/2007

3 DNA RNA RNA AllPrep DNA Spin Column DNA RNeasy MinElute Spin Column RNA 45 µg RNA Buffer RLT Plus AllPrep DNA RNeasy MinElute Spin Column Table 2 Handbook 14 RNA 5 x 10 5 RNA DNA AllPrep DNA Spin Column RNA RNeasy MinElute Spin Column HeLa 4 AllPrep DNA/RNA Micro Kit Important Notes Handbook 12 RNA Appendix A Handbook 44 TissueRuptor TissueRuptor User Manual TissueRuptor Handbook RNA 3,000 5,000 x g 5 RNase 4 AllPrep DNA/RNA Micro 07/2007 3

4 4. HeLa cm 2 * x x x x x mm 8 1 x ml 25 3 x 10 6 * HeLa 15 µm µm RNeasy MinElute Spin Column 5 x 10 5 AllPrep DNA Mini Spin Column RNAprotect Cell Reagent 5,000 x g 5 2 Buffer RLT Plus Buffer RW1 Buffer AW1 Safety information Handbook AllPrep DNA/RNA Micro 07/2007

5 RNase RNA β-me β-mercaptoethanol Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus Buffer RLT 1 ml 20 µl 2 M dithiothreitol DTT2 M DTT DTT Buffer RLT Plus RNA Handbook 17 Carrier RNA RNA 310 µg 1 ml RNase 20 RNA 310 µg/ml = 310 ng/µl10 4 ng/µl 5 µl RNA 34 µl Buffer RLT Plus 6 µl 54 µl Buffer RLT Plus 4 ng/µl 5 µl 3 oligo-dt RNA RNA Buffer RPE Buffer AW1 Buffer AW ml ml RNase Buffer RLT Plus DNA Buffer EB 70 AllPrep DNA/RNA Micro 07/2007 5

6 1. 1a1b 1a. 5 x x g 5 2 1b. 5 x cm 2 PBS PBS PBS RNase 300 x g Buffer RLT Plus 350 µl Buffer RLT Plus 3 1 x µl Buffer RLT Plus 6 AllPrep DNA/RNA Micro 07/2007

7 350 µl Buffer RLT Plus 3 1 x µl Buffer RLT Plus 3. 3a 3b3c Handbook 15 Disrupting and homogenizing starting material 1 x µl Buffer RLT Plus 1.5 ml Buffer RLT Plus 350 µl ng RNA 4 ng/µl 5 µl RNA RNA AllPrep Spin Column RNeasy MinElute Spin Column TissueRuptor QIAshredder 3a. 2 ml QIAshredder Spin Column 2 4 3b. TissueRuptor 30 TissueRuptor 4 TissueRuptor 3c. RNase 20-G 0.9 mm ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm30 AllPrep DNA/RNA Micro 07/2007 7

8 5. AllPrep DNA Spin Column 2 ml DNA RNA mirna small RNA RNA 6 12 Handbook 51 Appendix D D1 D6 AllPrep DNA Spin Column 4 RNA µl 7 DNA µl RNA 7. 2 ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * Handbook 53 Appendix E E1 E µl Buffer RW1 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm15 * 9 RNeasy MinElute Spin Column * Buffer RLT Plus Buffer RW1 Safety information Handbook 7 8 AllPrep DNA/RNA Micro 07/2007

9 9. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm Buffer RPE Buffer RPE µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm RNase 80 RNeasy MinElute Spin Column 11. RNeasy MinElute Spin Column2 ml 5 RNA 12. RNeasy MinElute Spin Column1.5 ml RNase 14 µl 1RNA RNA 10 µl RNase µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR RNA WTA QuantiTect Whole Transcriptome Kit AllPrep DNA/RNA Micro 07/2007 9

10 DNA AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm15 * 14 Buffer AW1 Buffer AW µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column AllPrep DNA Spin Column1.5 ml 50 µl Buffer EB ,000 x g 10,000 rpm1dna DNA DNA 1.5 ml 2 DNA 1 2 DNA 15 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR DNA QIAGEN * Buffer AW1 Safety information Handbook 7 10 AllPrep DNA/RNA Micro 07/2007

11 DNA RNA RNA RNA 19 5 mg RNAlater Allprotect 2 3 mg AllPrep DNA Spin Column DNA RNeasy MinElute Spin Column RNA Buffer RLT Plus DNA RNA Table 2 Handbook 14 RNA DNA RNA DNA DNA RNA RNeasy MinElute Spin Column DNase Handbook 54 Appendix F RNA DNA RNA DNeasy Blood & Tissue Kit RNeasy Fibrous Tissue Mini Kit Handbook 56 ordering information RNA DNA AllPrep DNA Spin Column RNA RNeasy MinElute Spin Column 1.5 mm 3.4 mm mg AllPrep DNA/RNA Micro Kit Important Notes Handbook 12 RNA Appendix A Handbook 44 TissueRuptor TissueRuptor User Manual TissueRuptor Handbook AllPrep DNA/RNA Micro 07/

12 TissueLyser TissueLyser Handbook RNAlater RNA Stabilization Reagent RNAlater Handbook Allprotect Tissue Reagent Allprotect Tissue Reagent Handbook RNAlater 4 Allprotect RNAlater/Allprotect Buffer RLT Plus RNA Buffer RLT Plus Buffer RW1 Buffer AW1 Safety information Handbook β- β-me Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus Buffer RLT Plus 1 ml 20 µl 2 M dithiothreitol DTT2 M DTT DTT Buffer RLT Plus 1 2 µg RNA Handbook 17 Carrier RNA RNA 310 µg 1 ml RNase 20 RNA 310 µg/ml = 310 ng/µl10 4 ng/µl 5 µl RNA 34 µl Buffer RLT Plus 6 µl 54 µl Buffer RLT Plus 4 ng/µl 5 µl 12 AllPrep DNA/RNA Micro 07/2007

13 2 oligo-dt RNA RNA Buffer RPE Buffer AW1 Buffer AW ml ml RNase Buffer RLT Plus DNA Buffer EB mg 2 RNAlater Allprotect RNAlater Allprotect RNA RNAlater RNA Stabilization Reagent Allprotect Reagent 80 RNAlater Allprotect Reagent 2 RNA RNA RNAlater Reagent DNA RNA Allprotect Tissue Reagent RNA AllPrep DNA/RNA Micro 07/

14 2. 2a 2b 2c Buffer RLT Plus 5 mg Handbook 15 Disrupting and homogenizing starting material β-me DTT Buffer RLT Plus 2 µg 20 ng RNA 4 ng/µl 5 µl RNA RNAlater Allprotect Reagent RNA AllPrep Spin Column RNeasy MinElute Spin Column TissueRuptor TissueLyser DNA 2a. TissueRuptor 350 µl Buffer RLT Plus TissueRuptor TissueRuptor 30 3 TissueRuptor 2 3 2b. TissueLyser 5 mm 1 2 ml 14 AllPrep DNA/RNA Micro 07/2007

15 350 µl Buffer RLT Plus TissueLyser Adapter Set 2 x 24 TissueLyser 20 Hz 2 TissueLyser 20 Hz2 3 2c. QIAshredder RNase 2 ml 350 µl Buffer RLT Plus 2 ml QIAshredder Spin Column 2 RNase 20-G ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm 30 3 AllPrep DNA/RNA Micro 07/

16 4. AllPrep DNA Spin Column2 ml DNA RNA AllPrep DNA Spin Column 4 RNA µl 6 DNA µl RNA ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * Handbook 53 Appendix E E1 E µl Buffer RW1 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm15 * 8 RNeasy MinElute Spin Column DNA RNA RNA 7 F1 F4 Handbook 54 Appendix F DNase * Buffer RLT Plus Buffer RW1 Safety information Handbook 7 16 AllPrep DNA/RNA Micro 07/2007

17 8. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm 15 9 Buffer RPE Buffer RPE µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm RNase 80 RNeasy MinElute Spin Column 10. RNeasy MinElute Spin Column2 ml 5 RNA 11. RNeasy MinElute Spin Column1.5 ml RNase 14 µl 1RNA RNA 10 µl RNase µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR RNA WTA QuantiTect Whole Transcriptome Kit AllPrep DNA/RNA Micro 07/

18 DNA AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm 15 * 13 Buffer AW1 Buffer AW µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column AllPrep DNA Spin Column1.5 ml Buffer EB µl ,000 x g 10,000 rpm1dna DNA DNA 1.5 ml 2 DNA 1 2 DNA 14 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR DNA QIAGEN * Buffer AW1 Safety information Handbook 7 18 AllPrep DNA/RNA Micro 07/2007

19 DNA RNA DNA RNA RNA RNeasy FFPE Kit RNA Leica P.A.L.M. Microlaser Technologies AllPrep DNA/RNA Micro Kit Important Notes Handbook 12 RNA Appendix A Handbook 44 RNA RNA RNA Buffer RLT Plus Buffer RW1 Buffer AW1 Safety information Handbook AllPrep DNA/RNA Micro 07/

20 β- β-me Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus Buffer RLT Plus 1 ml 20 µl 2 M dithiothreitol DTT2 M DTT DTT Buffer RLT Plus RNA Handbook 17 Carrier RNA RNA 310 µg 1 ml RNase 20 RNA 310 µg/ml = 310 ng/µl10 4 ng/µl 5 µl RNA 34 µl Buffer RLT Plus 6 µl 54 µl Buffer RLT Plus 4 ng/µl 5 µl 2 oligo-dt RNA RNA Buffer RPE Buffer AW1 Buffer AW ml ml RNase Buffer RLT Plus DNA Buffer EB Buffer RLT Plus 65 µl Leica 300 µl β-me DTT Buffer RLT Plus ml Buffer RLT Plus 350 µl 20 AllPrep DNA/RNA Micro 07/2007

21 ng RNA 4 ng/µl 5 µl RNA ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm AllPrep DNA Spin Column2 ml DNA RNA AllPrep DNA Spin Column 4 RNA µl 7 DNA µl 7. 2 ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * µl Buffer RW1 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm15 * 9 RNeasy MinElute Spin Column * Buffer RLT Plus Buffer RW1 Safety information Handbook 7 AllPrep DNA/RNA Micro 07/

22 9. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm Buffer RPE Buffer RPE µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm RNase 80 RNeasy MinElute Spin Column 11. RNeasy MinElute Spin Column2 ml 5 RNA 12. RNeasy MinElute Spin Column1.5 ml RNase 14 µl 1RNA RNA 10 µl RNase µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR RNA WTA QuantiTect Whole Transcriptome Kit 22 AllPrep DNA/RNA Micro 07/2007

23 DNA AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm 15 * 14 Buffer AW1 Buffer AW µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column AllPrep DNA Spin Column1.5 ml 50 µl Buffer EB ,000 x g 10,000 rpm2dna DNA DNA 1.5 ml 2 DNA 1 2 DNA 15 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR DNA QIAGEN * Buffer AW1 Safety information Handbook 7 AllPrep DNA/RNA Micro 07/

24 AllPrep DNARNeasy MinElute Spin Column a b c a b c RNA RNeasy MinElute Spin Column d DNA AllPrep DNA Spin Column Disrupting and homogenizing starting materials Handbook 15 Handbook AllPrep DNA Spin Column 37 Disrupting and homogenizing starting materials Handbook 15 RNA RNase RNeasy MinElute Spin Column 10 DNA Buffer EB AllPrep DNA Spin Column AllPrep DNA/RNA Micro 07/2007

25 e f DNA RNA a AllPrep DNA Spin Column b ph DNA RNA a AllPrep DNA Spin Column DNA RNA DNA b c DNA 80 RNeasy MinElute Spin Column 5 RNeasy MinElute Spin Column 3 AllPrep DNA Spin Column ph 7 AllPrep DNA Spin Column DNA DNA DNA AllPrep DNA Spin Column RNeasy MinElut Supin Column DNase Handbook 54 Appendix F AllPrep DNA/RNA Micro 07/

26 RNAA 260 /A 280 A 260 /A 280 RNA RNA a b RNase DNA RNase 10 mm Tris Cl* ph 7.5 Handbook 46 Appendix B Allprotect Tissue Reagent RNAlater RNA Stabilization Reagent 70 AllPrep DNA/RNA Appendix A Handbook 44 Handling and storage of starting material Handbook 14 AllPrep RNase RNase AllPrep DNA/RNA RNase RNA Handbook 44 Appendix A DNA kb DNA QIAshredder * MSDS material safety data sheet 26 AllPrep DNA/RNA Micro 07/2007

27 a b AllPrep DNA Spin Column 30 µl Buffer EB RNeasy 12 µl RNase 80 RNeasy MinElute Spin Column 5 RNeasy MinElute Spin Column AllPrep DNA/RNA Micro 07/

28 QIAGEN, AllPrep, DNeasy, MinElute, QuantiTect, RNAprotect, RNeasy (QIAGEN Group); Leica (Leica Microsystems GmbH). RNAlater is a trademark of AMBION, Inc., Austin, Texas and is covered by various U.S. and foreign patents. QIAGEN 2007 QIAGEN, all rights reserved Forefront Tower II Tel: Fax: /2007 Sample & Assay Technologies

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