PAXgene Tissue DNA Kitプロトコールとトラブルシューティング(2301605 06/2009)

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Transcription:

January 2009 PAXgene Tissue DNA Kit PAXgene Tissue Containers DNA PAXgene Tissue Container

PAXgene DNA 3 PAXgene DNA 6 PAXgene DNA 9 13 2 PAXgene Tissue DNA Kit 01/2009

PAXgene DNA DNA PAXgene Tissue Containers 2 5 PAXgene Tissue Container Product Circular 5 10 µm 100 mm 2 DNA DNAPAXgene DNA Spin Column DNA 15 25 PAXgene Tissue Container Product Circular Buffer TD1 Buffer TD2 56 Buffer TD3 Buffer TD4 96 100 purity grade p.a. 9 56 PAXgene Tissue DNA Kit 01/2009 3

1. 5 10 µm 2 5 2 3 2. 1.5 ml 3. 650 µl20 3 15 25 4. 650 µl 96 100 purity grade p.a. 20 5. 520,000 x g 20,000 x g 6. Proteinase KPAXgene Tissue DNA 7. 15 2537 10 8. 180 µl Buffer TD1 20 µl Proteinase K 15 9. 1,400 rpm 56 1 11 80 10. RNA DNA4 µl RNase A 100 mg/ml 15 25 2 11 11. 1,400 rpm 80 60 12. 1.5 ml 4 PAXgene Tissue DNA Kit 01/2009

13. 200 µl Buffer TD2 15 Buffer TD2 RNA Handbook 13 Carrier RNA 1 µg RNA 200 µl Buffer TD2 RNA Buffer TD2 Buffer TD5 Buffer TD2 14. 200 µl 96 100 PAXgene Tissue DNA 15. 1.5 ml 16. 2 mlpaxgene DNA Spin Column 6,000 x g 1 2 ml * 17. PAXgene DNA Spin Column 500 µl Buffer TD3 6,000 x g 12 ml * 18. PAXgene DNA Spin Column 500 µl Buffer TD4 6,000 x g 1 PAXgene 2 ml 19. 3 20,000 x g 20. PAXgene DNA Spin Column 1.5 ml 20 200 µl Buffer TD5 PAXgene DNA Spin Column 1 20,000 x g DNA Buffer TD5 PAXgene DNA Spin Column 15 255 DNA * Buffer TD2 Buffer TD3 Safety information Handbook 6 PAXgene Tissue DNA Kit 01/2009 5

PAXgene DNA DNA PAXgene Tissue Containers 20 mg DNA 10 mg DNA PAXgene Tissue Container 20 mg PAXgene DNA Spin Column DNA 2 mm 8 mm 3 8 12 mg DNAPAXgene DNA Spin Column DNA 15 25 PAXgene Tissue Container Product Circular Buffer TD1 Buffer TD2 56 6 PAXgene Tissue DNA Kit 01/2009

Buffer TD3 Buffer TD4 96 100 purity grade p.a. 5 56 1. PAXgene Tissue Container 20 mg10 mg 2 ml 180 µl Buffer TD1 2. 2 ml 5 mm 1 TissueLyser Adapter Set 2 x 24 TissueRuptor DNA 3. TissueLyser 15 Hz 20 DNA 4. 1.5 ml 5. 20 µl Proteome K 1,400 rpm 56 1 8 70 TissueLyser 3 6. RNA DNA4 µl RNase A 100 mg/ml 15 25 2 7. 200 µl Buffer TD2 15 Buffer TD2 8. 70 10 PAXgene Tissue DNA Kit 01/2009 7

9. 200 µl 96 100 PAXgene Tissue DNA 10. 11. 2 mlpaxgene DNA Spin Column 6,000 x g 1 2 ml * 12. PAXgene DNA Spin Column 500 µl Buffer TD3 6,000 x g 12 ml * 13. PAXgene DNA Spin Column 500 µl Buffer TD4 6,000 x g 12 ml 14. 3 20,000 x g 15. PAXgene DNA Spin Column 1.5 ml 50 200 µl Buffer TD5 PAXgene DNA Spin Column 1 20,000 x g DNA Buffer TD5 PAXgene DNA Spin Column 15 255 DNA 16. 15 * Buffer TD2 Buffer TD3 Safety information Handbook 6 8 PAXgene Tissue DNA Kit 01/2009

PAXgene DNA DNA PAXgene Tissue Containers 20 mg PAXgene Tissue Container Product CircularDNA 10 mg DNA 20 mg PAXgene DNA Spin Column DNA 2 mm 8 mm 3 8 12 mg DNAPAXgene DNA Spin Column DNA 15 25 PAXgene Tissue Container Product Circular Buffer TD1 Buffer TD2 56 PAXgene Tissue DNA Kit 01/2009 9

Buffer TD3 Buffer TD4 96 100 purity grade p.a 15 56 1. 20 mg10 mg 2. 2 ml 3. 1 ml203 15 25 4. 320,000 x g 20,000 x g 5. 6. 1 ml 96 100 purity grade p.a. 20 7. 320,000 x g 8. 9. 15 2537 10 10. 180 µl Buffer TD1 11. 2 ml 5 mm 1 TissueLyser Adapter Set 2 x 24 TissueRuptor DNA 12. TissueLyser 15 Hz 20 DNA 13. 1.5 ml 10 PAXgene Tissue DNA Kit 01/2009

14. 20 µl Proteinase K 15 15. 1,400 rpm 56 1 17 80 16. RNA DNA4 µl RNase A 100 mg/ml 15 25 2 17. 1,400 rpm 80 60 18. 1.5 ml 19. 200 µl Buffer TD2 15 Buffer TD2 20. 200 µl 96 100 PAXgene Tissue DNA 21. 1.5 ml 22. 2 mlpaxgene DNA Spin Column 6,000 x g 1 2 ml * 23. PAXgene DNA Spin Column 500 µl Buffer TD3 6,000 x g 12 ml * 24. PAXgene DNA Spin Column 500 µl Buffer TD4 6,000 x g 12 ml * Buffer TD2 Buffer TD3 Safety information Handbook 6 PAXgene Tissue DNA Kit 01/2009 11

25. 3 20,000 x g 26. PAXgene DNA Spin Column 1.5 ml 50 200 µl Buffer TD5 PAXgene DNA Spin Column 1 20,000 x g DNA Buffer TD5 PAXgene DNA Spin Column 15 25 5 DNA 12 PAXgene Tissue DNA Kit 01/2009

a b Buffer TD2 c Buffer TD3 Buffer TD4 d DNA PAXgene DNA Spin Column A 260 /A 280 Buffer TD2 15 PAXgene DNA Spin Column Buffer TD3 Buffer TD4 3 6 9 DNA 56 Proteinase K g DNA A 260 /A 280 10 mm Tris Cl ph 7.5* Handbook 28 Appendix: Determination of Yield, Purity, and Length of DNA DNA A 260 /A 280 RNA RNase * MSDS material safety data sheet PAXgene Tissue DNA Kit 01/2009 13

DNA a b c DNA DNA a DNA b DNA TissueLyser Buffer TD4 15 25 Buffer TD3 Buffer TD4 Buffer TD4 PAXgene DNA Spin Column 3 PAXgene DNA Spin Column PAXgene DNA Spin Column 1 PCR 100 ng DNA 35 PCR PAXgene 2 8 15 30 TissueLyser 15 Hz 20 14 PAXgene Tissue DNA Kit 01/2009

Trademarks: PAXgene, PreAnalytiX (PreAnalytiX GmbH.); QIAGEN, TissueRuptor (QIAGEN Group). For updated license terms, see www.preanalytix.com. 2009 PreAnalytiX GmbH, all rights reserved. 104-0054 3-13-1 Forefront Tower II Tel:03-6890-7300 Fax:03-5547-0818 E-mail:techservice-jp@qiagen.com www.qiagen.co.jp PreAnalytiX Distributors PreAnalytiX products are manufactured for PreAnalytiX by QIAGEN or BD and are distributed for PreAnalytiX by QIAGEN or BD. Products cannot be ordered at PreAnalytiX GmbH.

2301605 06/2009