July 2007 AllPrep DNA/RNA Micro DNA RNA 5 x 10 5 5 mg Sample & Assay Technologies
DNA RNA 3 DNA RNA 11 DNA RNA 19 24 2 AllPrep DNA/RNA Micro 07/2007
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 70 2 3 70 37 RNA 3,000 5,000 x g 5 RNase 4 AllPrep DNA/RNA Micro 07/2007 3
4. HeLa cm 2 * 96 0.32 0.6 4 5 x 10 4 48 1 1 x 10 5 24 2 2.5 x 10 5 12 4 5 x 10 5 6 9.5 1 x 10 6 35 mm 8 1 x 10 6 40 50 ml 25 3 x 10 6 * HeLa 15 µm 10 30 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 7 15 25 20 25 20 4 AllPrep DNA/RNA Micro 07/2007
RNase RNA β-me β-mercaptoethanol Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus 15 25 1 Buffer RLT 1 ml 20 µl 2 M dithiothreitol DTT2 M DTT DTT Buffer RLT Plus 1 500 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 AW2 96 100 24 ml 96 100 6 ml RNase 80 70 96 100 Buffer RLT Plus DNA Buffer EB 70 AllPrep DNA/RNA Micro 07/2007 5
1. 1a1b 1a. 5 x 10 5 300 x g 5 2 1b. 5 x 10 5 10 cm 2 PBS PBS 0.10 0.25 PBS RNase 300 x g 5 2 2. Buffer RLT Plus 350 µl Buffer RLT Plus 3 1 x 10 5 75 µl Buffer RLT Plus 6 AllPrep DNA/RNA Micro 07/2007
350 µl Buffer RLT Plus 3 1 x 10 5 75 µl Buffer RLT Plus 3. 3a 3b3c Handbook 15 Disrupting and homogenizing starting material 1 x 10 5 1 4 2 75 µl Buffer RLT Plus 1.5 ml Buffer RLT Plus 350 µl 1 4 500 20 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 54 4. 2 ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm30 AllPrep DNA/RNA Micro 07/2007 7
5. AllPrep DNA Spin Column 2 ml 15 25 13 16 DNA 4 6 12 RNA mirna small RNA RNA 6 12 Handbook 51 Appendix D D1 D6 AllPrep DNA Spin Column 4 RNA 6. 5 70 350 µl 7 DNA 70 350 µl RNA 7. 2 ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * Handbook 53 Appendix E E1 E5 8 8. 700 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. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm 15 10 Buffer RPE Buffer RPE 10. 500 µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 2 96 100 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 20 10 µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR www.qiagen.com/pcr RNA WTA QuantiTect Whole Transcriptome Kit www.qiagen.com/goto/wta AllPrep DNA/RNA Micro 07/2007 9
DNA 13. 5 AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm15 * 14 Buffer AW1 Buffer AW1 14. 500 µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column 1 15. AllPrep DNA Spin Column1.5 ml 50 µl Buffer EB 70 15 25 2 8,000 x g 10,000 rpm1dna 16. 15 DNA DNA 1.5 ml 2 DNA 1 2 DNA 15 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR www.qiagen.com/pcr DNA QIAGEN www.qiagen.com/wga * Buffer AW1 Safety information Handbook 7 10 AllPrep DNA/RNA Micro 07/2007
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 5 70 50 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 3 3.5 4.5 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 07/2007 11
TissueLyser TissueLyser Handbook RNAlater RNA Stabilization Reagent RNAlater Handbook Allprotect Tissue Reagent Allprotect Tissue Reagent Handbook 37 1 15 25 7 2 8 RNAlater 4 Allprotect 6 20 80 RNAlater/Allprotect 70 70 Buffer RLT Plus 2 70 3 37 RNA Buffer RLT Plus Buffer RW1 Buffer AW1 Safety information Handbook 7 15 25 20 25 20 β- β-me Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus 15 25 1 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
2 oligo-dt RNA RNA Buffer RPE Buffer AW1 Buffer AW2 96 100 24 ml 96 100 6 ml RNase 80 70 96 100 Buffer RLT Plus DNA Buffer EB 70 1. 5 mg 2 RNAlater Allprotect RNAlater Allprotect RNA 15 25 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 07/2007 13
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
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 53 3. 3 2 ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm 30 3 AllPrep DNA/RNA Micro 07/2007 15
4. AllPrep DNA Spin Column2 ml 12 15 DNA 15 25 4 5 11 RNA AllPrep DNA Spin Column 4 RNA 5. 4 70 350 µl 6 DNA 70 350 µl RNA 70 50 6. 2 ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * Handbook 53 Appendix E E1 E5 7 7. 700 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
8. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm 15 9 Buffer RPE Buffer RPE 9. 500 µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 2 96 100 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 20 10 µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR www.qiagen.com/pcr RNA WTA QuantiTect Whole Transcriptome Kit www.qiagen.com/goto/wta AllPrep DNA/RNA Micro 07/2007 17
DNA 12. 4 AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm 15 * 13 Buffer AW1 Buffer AW1 13. 500 µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column 1 14. AllPrep DNA Spin Column1.5 ml Buffer EB 70 50 µl 15 25 2 8,000 x g 10,000 rpm1dna 15. 14 DNA DNA 1.5 ml 2 DNA 1 2 DNA 14 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR www.qiagen.com/pcr DNA QIAGEN www.qiagen.com/wga * Buffer AW1 Safety information Handbook 7 18 AllPrep DNA/RNA Micro 07/2007
DNA RNA DNA RNA RNA RNeasy FFPE Kit RNA Leica www.leica-microsystems.co.jp P.A.L.M. Microlaser Technologies www.palm-mikrolaser.com AllPrep DNA/RNA Micro Kit Important Notes Handbook 12 RNA Appendix A Handbook 44 RNA RNA 3 70 4 37 RNA Buffer RLT Plus Buffer RW1 Buffer AW1 Safety information Handbook 7 15 25 20 25 20 AllPrep DNA/RNA Micro 07/2007 19
β- β-me Buffer RLT Plus 1 ml Buffer RLT Plus β-me 10 µl β-me Buffer RLT Plus 15 25 1 Buffer RLT Plus 1 ml 20 µl 2 M dithiothreitol DTT2 M DTT DTT Buffer RLT Plus 1 500 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 AW2 96 100 24 ml 96 100 6 ml RNase 80 70 96 100 Buffer RLT Plus DNA Buffer EB 70 1. Buffer RLT Plus 65 µl Leica 300 µl β-me DTT Buffer RLT Plus 2. 1.5 2 ml Buffer RLT Plus 350 µl 20 AllPrep DNA/RNA Micro 07/2007
500 20 ng RNA 4 ng/µl 5 µl RNA 3. 30 4. 2 ml AllPrep DNA Spin Column 8,000 x g 10,000 rpm 30 5. AllPrep DNA Spin Column2 ml DNA 13 1615 25 4 6 12 RNA AllPrep DNA Spin Column 4 RNA 6. 5 70 350 µl 7 DNA 70 350 µl 7. 2 ml RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 15 * 8 8. 700 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 07/2007 21
9. RNeasy MinElute Spin Column Buffer RPE 500 µl 8,000 x g 10,000 rpm 15 10 Buffer RPE Buffer RPE 10. 500 µl 80 RNeasy MinElute Spin Column 8,000 x g 10,000 rpm 2 96 100 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 20 10 µl RNase RNA RNeasy MinElute Spin Column 2 µl 14 µl RNase 12 µl RNA RT-PCR RT-PCR www.qiagen.com/pcr RNA WTA QuantiTect Whole Transcriptome Kit www.qiagen.com/goto/wta 22 AllPrep DNA/RNA Micro 07/2007
DNA 13. 5 AllPrep DNA Spin Column 500 µl Buffer AW1 8,000 x g 10,000 rpm 15 * 14 Buffer AW1 Buffer AW1 14. 500 µl Buffer AW2 AllPrep DNA Spin Column 2 Buffer AW2 Buffer AW2 DNA AllPrep DNA Spin Column 1 15. AllPrep DNA Spin Column1.5 ml 50 µl Buffer EB 70 15 25 1 8,000 x g 10,000 rpm2dna 16. 15 DNA DNA 1.5 ml 2 DNA 1 2 DNA 15 DNA 30 µl Buffer EB 2 DNA DNA PCR PCR www.qiagen.com/pcr DNA QIAGEN www.qiagen.com/wga * Buffer AW1 Safety information Handbook 7 AllPrep DNA/RNA Micro 07/2007 23
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 12 20 25 20 20 25 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 10 24 AllPrep DNA/RNA Micro 07/2007
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 07/2007 25
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 15 30 kb DNA QIAshredder * MSDS material safety data sheet 26 AllPrep DNA/RNA Micro 07/2007
a 20 30 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 07/2007 27
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. 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 2301246 07/2007 Sample & Assay Technologies