HB QIAamp Circulating Nucleic Acid (05/2014)



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Transcription:

January 2011 QIAamp Circulating Nucleic Acid DNA RNA mirna Sample & Assay Technologies

1 ml 2 ml 3 ml 3 4 ml 5 ml 7 1 ml 2 ml 3 ml 11 4 ml 15 1 ml 2 ml 3 ml microrna 19 22 2 QIAamp Circulating Nucleic Acid 01/2011

1 ml 2 ml 3 ml 1 ml 2 ml 3 ml DNA RNA 4 ml 5 ml 7 4 ml 5 ml DNA RNA 1 ml 2 ml 3 ml 11 4 ml 15 RNA mirna 1 ml 2 ml 3 ml microrna 19 1 ml 2 ml 3 ml 15 25 15 25 <1 ml <2 ml <3 ml PBS phosphate-buffered saline 1 ml 2 ml 3 ml QIAvac 24 Plus Handbook 18 21 4 50 ml 60 14 2 ml 56 15 Buffer AVE Buffer ACB Buffer ACW1 Buffer ACW2 Handbook 14 Handbook 14 15 2 Buffer AVE RNA Buffer ACL QIAamp Circulating Nucleic Acid 01/2011 3

2. 1 ml 2 ml 3 ml Buffer ACL RNA Buffer AVE Buffer ACL ml Buffer AVE RNA µl 1 0.9 1.8 2.6 5.6 2 1.8 3.5 5.3 11.3 3 2.6 5.3 7.9 16.9 4 3.5 7.0 10.6 22.5 5 4.4 8.8 13.2 28.1 6 5.3 10.6 15.8 33.8 7 6.2 12.3 18.5 39.4 8 7.0 14.1 21.1 45.0 9 7.9 15.8 23.8 50.6 10 8.8 17.6 26.4 56.3 11 9.7 19.4 29.0 61.9 12 10.6 21.1 31.7 67.5 13 11.4 22.9 34.3 73.1 14 12.3 24.6 37.0 78.8 15 13.2 26.4 39.6 84.4 16 14.1 28.2 42.2 90.0 17 15.0 29.9 44.9 95.6 18 15.8 31.7 47.5 101.3 19 16.7 33.4 50.2 106.9 20 17.6 35.2 52.8 112.5 21 18.5 37.0 55.4 118.1 22 19.4 38.7 58.1 123.8 23 20.2 40.5 60.7 129.4 24 21.1 42.2 63.4 135.0 4 QIAamp Circulating Nucleic Acid 01/2011

1. 50 ml 100 µl 200 µl 300 µl QIAGEN Proteinase K 2. 50 ml 1 ml 2 ml 3 ml 3. 0.8 ml 1.6 ml 2.4 ml Buffer ACL 1.0 µg RNA 30 Buffer ACL 4 4. 60 30 5. 6. 1.8 ml 3.6 ml 5.4 ml Buffer ACB 15 30 7. Buffer ACB 5 8. QIAvac 24 Plus VacConnector QIAamp Mini Column 20 ml Tube Extender QIAamp Mini Column Tube Extender QIAamp Mini Column 13 9. 7 Buffer ACB QIAamp Mini Column Tube Extender 0 mbar Tube Extender 3 ml 11 ml QIAamp Mini 10 Vacuum Regulator QIAvac Connecting System Tube Extender QIAamp Mini Column 10. 600 µl Buffer ACW1 QIAamp Mini Column Buffer ACW1 QIAamp Mini Column 0 mbar QIAamp Circulating Nucleic Acid 01/2011 5

11. 750 µl Buffer ACW2 QIAamp Mini Column Buffer ACW2 QIAamp Mini Column 0 mbar 12. 750 µl 96 100 QIAamp Mini Column 0 mbar 13. QIAamp Mini Column VacConnector QIAamp Mini Column 2 ml 8 20,000 x g 14,000 rpm 3 14. QIAamp Mini Column 2 ml 56 10 15. QIAamp Mini Column 1.5 ml 14 2 ml 20 150 µl Buffer AVE QIAamp Mini 15 25 3 Buffer AVE 15 25 50 µl QIAamp Mini Column 5 µl 16. 20,000 x g 14,000 rpm 1 6 QIAamp Circulating Nucleic Acid 01/2011

4 ml 5 ml 4 ml 5 ml DNA RNA 1 ml 2 ml 3 ml 3 1 ml 2 ml 3 ml DNA RNA 1 ml 2 ml 3 ml 11 4 ml 15 RNA mirna 1 ml 2 ml 3 ml microrna 19 4 ml 5 ml 15 25 15 25 <4 ml <5 ml PBS phosphate-buffered saline 4 ml 5 ml QIAvac 24 Plus Handbook 18 21 4 50 ml 60 14 2 ml 56 15 Buffer AVE Buffer ACB Buffer ACW1 Buffer ACW2 Handbook 14 Handbook 14 15 3 Buffer AVE RNA Buffer ACL QIAamp Circulating Nucleic Acid 01/2011 7

3. 4 ml 5 ml Buffer ACL RNA Buffer AVE Buffer ACL ml Buffer AVE RNA µl 1 3.5 4.4 5.6 2 7.0 8.8 11.3 3 10.6 13.2 16.9 4 14.1 17.6 22.5 5 17.6 22.0 28.1 6 21.1 26.4 33.8 7 24.6 30.8 39.4 8 28.2 35.2 45.0 9 31.7 39.6 50.6 10 35.2 44.0 56.3 11 38.7 48.4 61.9 12 42.2 52.8 67.5 13 45.8 57.2 73.1 14 49.3 61.6 78.8 15 52.8 66.0 84.4 16 56.3 70.4 90.0 17 59.8 74.8 95.6 18 63.4 79.2 101.3 19 66.9 83.6 106.9 20 70.4 88.0 112.5 21 73.9 92.4 118.1 22 77.4 96.8 123.8 23 81.0 101.2 129.4 24 84.5 105.6 135.0 8 QIAamp Circulating Nucleic Acid 01/2011

1. 50 ml 400 µl 500 µl QIAGEN Proteinase K 2. 4 ml 5 ml 3. 3.2 ml 4.0 ml Buffer ACL 1.0 µg RNA 30 Buffer ACL 4 4. 60 30 5. 6. 7.2 ml 9 ml Buffer ACB 15 30 7. Buffer ACB 5 8. QIAvac 24 Plus VacConnector QIAamp Mini Column 20 ml Tube Extender QIAamp Mini Column Tube Extender QIAamp Mini Column 13 9. 7 Buffer ACB QIAamp Mini Column Tube Extender 0 mbar Tube Extender 5 ml 20 ml QIAamp Mini 15 Vacuum Regulator QIAvac Connecting System Tube Extender QIAamp Mini Column 10. 600 µl Buffer ACW1 QIAamp Mini Column Buffer ACW1 QIAamp Mini Column 0 mbar QIAamp Circulating Nucleic Acid 01/2011 9

11. 750 µl Buffer ACW2 QIAamp Mini Column Buffer ACW2 QIAamp Mini Column 0 mbar 12. 750 µl 96 100 QIAamp Mini Column 0 mbar 13. QIAamp Mini Column VacConnector QIAamp Mini Column 2 ml 8 20,000 x g 14,000 rpm 3 14. QIAamp Mini Column 2 ml 56 10 15. QIAamp Mini Column 1.5 ml 14 2 ml 20 150 µl Buffer AVE QIAamp Mini 15 25 3 Buffer AVE 15 25 50 µl QIAamp Mini Column 5 µl 16. 20,000 x g 14,000 rpm 1 10 QIAamp Circulating Nucleic Acid 01/2011

1 ml 2 ml 3 ml 1 ml 2 ml 3 ml DNA RNA 4 ml 4 ml 15 1 ml 2 ml 3 ml 3 4 ml 5 ml 7 mirna 1 ml 2 ml 3 ml microrna 19 1 ml 2 ml 3 ml 16,000 x g 10 15 25 15 25 <1 ml <2 ml <3 ml PBS phosphate-buffered saline 1 ml 2 ml 3 ml QIAvac 24 Plus Handbook 18 21 4 50 ml 60 14 2 ml 56 15 Buffer AVE Buffer ACB Buffer ACW1 Buffer ACW2 Handbook 14 Handbook 14 15 4 Buffer AVE RNA Buffer ACL QIAamp Circulating Nucleic Acid 01/2011 11

4. 1 ml 2 ml 3 ml Buffer ACL RNA Buffer AVE Buffer ACL ml Buffer AVE RNA µl 1 0.9 1.8 2.6 5.6 2 1.8 3.5 5.3 11.3 3 2.6 5.3 7.9 16.9 4 3.5 7.0 10.6 22.5 5 4.4 8.8 13.2 28.1 6 5.3 10.6 15.8 33.8 7 6.2 12.3 18.5 39.4 8 7.0 14.1 21.1 45.0 9 7.9 15.8 23.8 50.6 10 8.8 17.6 26.4 56.3 11 9.7 19.4 29.0 61.9 12 10.6 21.1 31.7 67.5 13 11.4 22.9 34.3 73.1 14 12.3 24.6 37.0 78.8 15 13.2 26.4 39.6 84.4 16 14.1 28.2 42.2 90.0 17 15.0 29.9 44.9 95.6 18 15.8 31.7 47.5 101.3 19 16.7 33.4 50.2 106.9 20 17.6 35.2 52.8 112.5 21 18.5 37.0 55.4 118.1 22 19.4 38.7 58.1 123.8 23 20.2 40.5 60.7 129.4 24 21.1 42.2 63.4 135.0 12 QIAamp Circulating Nucleic Acid 01/2011

1. 50 ml 125 µl 250 µl 375 µl QIAGEN Proteinase K 2. 50 ml 1 ml 2 ml 3 ml 3. 1 ml 2 ml 3 ml Buffer ACL 1.0 µg RNA 250 µl 500 µl 750 µl Buffer ATL 30 Buffer ATL Buffer ACL ATL 4 4. 60 30 5. 6. 3.6 ml 5.4 ml 7.2 ml Buffer ACB 15 30 7. Buffer ACB 5 8. QIAvac 24 Plus VacConnector QIAamp Mini Column 20 ml Tube Extender QIAamp Mini Column Tube Extender QIAamp Mini Column 13 9. 7 QIAamp Mini Column Tube Extender 0 mbar Tube Extender 4 ml 20 ml QIAamp Mini 15 Vacuum Regulator QIAvac Connecting System Tube Extender QIAamp Mini Column QIAamp Circulating Nucleic Acid 01/2011 13

10. 600 µl Buffer ACW1 QIAamp Mini Column Buffer ACW1 QIAamp Mini Column 0 mbar 11. 750 µl Buffer ACW2 QIAamp Mini Column Buffer ACW2 QIAamp Mini Column 0 mbar 12. 750 µl 96 100 QIAamp Mini Column QIAamp Mini Column 0 mbar 13. QIAamp Mini Column VacConnector QIAamp Mini Column 2 ml 8 20,000 x g 14,000 rpm 3 14. QIAamp Mini Column 2 ml 56 10 4 4 15. QIAamp Mini Column 1.5 ml 14 2 ml 20 150 µl Buffer AVE QIAamp Mini 15 25 3 16. 20,000 x g 14,000 rpm 1 14 QIAamp Circulating Nucleic Acid 01/2011

4 ml 4 ml DNA RNA 1 ml 2 ml 3 ml 1 ml 2 ml 3 ml 11 1 ml 2 ml 3 ml 3 4 ml 5 ml 7 mirna 1 ml 2 ml 3 ml microrna 19 16,000 x g 10 15 25 15 25 QIAvac 24 Plus Handbook 18 21 4 50 ml 60 14 2 ml 56 15 Buffer AVE Buffer ACB Buffer ACW1 Buffer ACW2 Handbook 14 Handbook 14 15 5 Buffer AVE RNA Buffer ACL QIAamp Circulating Nucleic Acid 01/2011 15

5. 4 ml Buffer ACL RNA Buffer AVE Buffer ACL ml Buffer AVE RNA µl 1 4.4 5.6 2 8.8 11.3 3 13.2 16.9 4 17.6 22.5 5 22.0 28.1 6 26.4 33.8 7 30.8 39.4 8 35.2 45.0 9 39.6 50.6 10 44.0 56.3 11 48.4 61.9 12 52.8 67.5 13 57.2 73.1 14 61.6 78.8 15 66.0 84.4 16 70.4 90.0 17 74.8 95.6 18 79.2 101.3 19 83.6 106.9 20 88.0 112.5 21 92.4 118.1 22 96.8 123.8 23 101.2 129.4 24 105.6 135.0 16 QIAamp Circulating Nucleic Acid 01/2011

1. 50 ml 500 µl QIAGEN Proteinase K 2. 50 ml 4 ml 3. 4 ml Buffer ACL RNA 1.0 ml Buffer ATL 30 Buffer ATL Buffer ACL ATL 4 4. 60 30 5. 6. 9.0 ml Buffer ACB 15 30 7. Buffer ACB 5 8. QIAvac 24 Plus VacConnector QIAamp Mini Column 20 ml Tube Extender QIAamp Mini Column Tube Extender QIAamp Mini Column 13 9. 7 QIAamp Mini Column Tube Extender 0 mbar Tube Extender 4 ml 20 ml QIAamp Mini 15 Vacuum Regulator QIAvac Connecting System Tube Extender QIAamp Mini Column QIAamp Circulating Nucleic Acid 01/2011 17

10. 600 µl Buffer ACW1 QIAamp Mini Column Buffer ACW1 QIAamp Mini Column 0 mbar 11. 750 µl Buffer ACW2 QIAamp Mini Column Buffer ACW2 QIAamp Mini Column 0 mbar 12. 750 µl 96 100 QIAamp Mini Column QIAamp Mini Column 0 mbar 13. QIAamp Mini Column VacConnector QIAamp Mini Column 2 ml 8 20,000 x g 14,000 rpm 3 14. QIAamp Mini Column 2 ml 56 10 4 4 15. QIAamp Mini Column 1.5 ml 14 2 ml 20 150 µl Buffer AVE QIAamp Mini 15 25 3 16. 20,000 x g 14,000 rpm 1 18 QIAamp Circulating Nucleic Acid 01/2011

1 ml 2 ml 3 ml microrna 1 ml 2 ml 3 ml mirna RNA DNA RNA 1 ml 2 ml 3 ml 3 4 ml 5 ml 7 DNA RNA 1 ml 2 ml 3 ml 11 4 ml 15 1 ml 2 ml 3 ml 16,000 x g 10 15 25 15 25 <1 ml <2 ml <3 ml PBS phosphate-buffered saline 1 ml 2 ml 3 ml QIAvac 24 Plus Handbook 18 21 4 50 ml 60 14 2 ml 56 15 Buffer AVE Buffer ACB Buffer ACW1 Buffer ACW2 Handbook 14 Handbook 14 15 4 Buffer AVE RNA Buffer ACL QIAamp Circulating Nucleic Acid 01/2011 19

1. 50 ml 130 µl 270 µl 400 µl QIAGEN Proteinase K 2. 50 ml 1 ml 2 ml 3 ml 3. 1.1 ml 2.1 ml 3.2 ml Buffer ACL RNA 0.33 ml 0.67 ml 1.0 ml Buffer ATL 30 Buffer ATL Buffer ACL ATL 4. 60 30 5. 6. 3.0 ml 6.0 ml 9.0 ml Buffer ACB 2.3 ml 4.7 ml 7.0 ml 15 30 7. Buffer ACB 5 8. QIAvac 24 Plus VacConnector QIAamp Mini Column 20 ml Tube Extender QIAamp Mini Column Tube Extender QIAamp Mini Column 13 9. QIAamp Mini Column Tube Extender 7 0 mbar Tube Extender 3 ml 23 ml QIAamp Mini 15 Vacuum Regulator QIAvac Connecting System Tube Extender QIAamp Mini Column 10. 600 µl Buffer ACW1 QIAamp Mini Column Buffer ACW1 QIAamp Mini Column 0 mbar 20 QIAamp Circulating Nucleic Acid 01/2011

11. 750 µl Buffer ACW2 QIAamp Mini Column Buffer ACW2 QIAamp Mini Column 0 mbar 12. 750 µl 96 100 QIAamp Mini Column QIAamp Mini Column 0 mbar 13. QIAamp Mini Column VacConnector QIAamp Mini Column 2 ml 8 20,000 x g 14,000 rpm 3 14. QIAamp Mini Column 2 ml 56 10 4 4 15. QIAamp Mini Column 1.5 ml 14 2 ml 20 150 µl Buffer AVE QIAamp Mini 15 25 3 16. 20,000 x g 14,000 rpm 1 QIAamp Circulating Nucleic Acid 01/2011 21

a EDTA b c 2 d DNA e Buffer ACL f Buffer ACL RNA g 96 100 h Buffer ACB i Buffer ACW1 Buffer ACW2 i Buffer ACW1 Buffer ACW2 70 EDTA DNA DNA DNA QIAGEN Proteinase K QIAGEN Proteinase K Buffer ACL RNA 10 Buffer ACL RNA 96 100 Buffer ACB Handbook 14 Buffer ACW1 ACW2 Handbook 14 Buffer ACW1 ACW2 96 100 Handbook 14 22 QIAamp Circulating Nucleic Acid 01/2011

DNA RNA a DNA b c d Taq DNA PCR e RNA 22 Buffer ACW2 PCR RNA RNA QIAamp Circulating Nucleic Acid 01/2011 23

a QIAamp Mini Column b VacValve Tube Extender Tube Extender/QIAamp Mini Column/ VacConnector/VacValve QIAvac 24 Plus QIAamp Mini Column Tube Extender 50 ml QIAamp Mini Column 2 ml 1 QIAamp Mini Column Tube Extender VacConnector VacValve 50 ml Tube Extender VacValve QIAamp Mini Column QIAamp Mini Column QIAamp Mini Column 2 16,000 x g 5 QIAamp Mini Column 5 µl 24 QIAamp Circulating Nucleic Acid 01/2011

b 800 900 mbar QIAvac VacValve VacValve VacConnector luer extension QIAamp Mini Column VacConnector VacValve Luer cap luer extension vacuum regulator valve QIAamp Circulating Nucleic Acid 01/2011 25

Memo 26 QIAamp Circulating Nucleic Acid 01/2011

Trademarks: QIAGEN, QIAamp (QIAGEN Group). QIAGEN 2011 QIAGEN, all rights reserved.

www.qiagen.co.jp 104-0054 3-13-1 Forefront Tower II Tel:03-6890-7300 Fax:03-5547-0818 E-mail:techservice-jp@qiagen.com 2301837 02/2011 Sample & Assay Technologies