QIAquick Spin プロトコールとトラブルシューティング ( /2008)

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Transcription:

March 2008 QIAquick Spin QIAquick PCR Purification Kit PCR 100 bp 10 kb QIAquick Nucleotide Removal Kit 17 40mer DNA 40 bp 10 kb QIAquick Gel Extraction Kit DNA 70 bp 10 kb Sample & Assay Technologies

QIAquick PCR Purification Kit 3 5 QIAquick Nucleotide Removal Kit 8 QIAquick Gel Extraction Kit 10 13 16 2 QIAquick Spin 03/2008

QIAquick PCR Purification Kit PCR Handbook 8 DNA PCR MinElute Reaction Cleanup Kit QIAquick 100 bp 10 kb 96 100 Buffer PE 17,900 x g 13,000 rpm ph indicator I Buffer PB 1 250 120 µl ph indicator I 30 ml Buffer PB 600 µl ph indicator I 150 ml Buffer PB ph indicator I Buffer PB ph 7.5 ph indicator I ph indicator I PCR ph indicator I Buffer PB 1. PCR 5 Buffer PB 100 µl PCR 500 µl Buffer PB 2. ph indicator I Buffer PB 3M ph 5.0 10 µl 3. 2 mlqiaquick 4. DNA QIAquick 30 60 5. QIAquick QIAquick Spin 03/2008 3

6. 750 µl Buffer PE QIAquick 30 60 7. QIAquick 1 Buffer PE 8. 1.5 mlqiaquick 9. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 7.0 8.5 QIAquick 1 DNA 30 µl QIAquick 1 1 DNA QIAquick 50 µl 48 µl 30 µl 28 µl ph ph 7.0 8.5 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 10. DNALoading Dye 5 DNA Loading dye 3 bromophenol blue xylene cyanol orange GDNA Table 2 Handbook 15 4 QIAquick Spin 03/2008

QIAquick PCR Purification Kit QIAquick QIAvac Luer Adaptors QIAvac 6S/QIAvac 24 Plus PCR Handbook 8 DNA PCR MinElute Reaction Cleanup Kit 100 bp 10 kb 96 100 Buffer PE ph indicator I Buffer PB 1 250 120 µl ph indicator I 30 ml Buffer PB600 µl ph indicator I 150 ml Buffer PB ph indicator I Buffer PB ph 7.5 ph indicator I ph indicator I PCR ph indicator I Buffer PB 1. PCR 5 Buffer PB 100 µl PCR 500 µl Buffer PB 2. ph indicator I Buffer PB 3M ph 5.0 10 µl 3. 3a 3b 3c QIAquick 3a. QIAvac 24 PlusHandbook 33Figure 7 QIAvac 24 Plus 24 QIAquick QIAvac 24 Plus QIAquick Spin 03/2008 5

3b. QIAvac 6S Handbook 34Figure 8 QIAvac 6S QIAvac QIAvac Luer Adaptor QIAvac 6S QIAvac QIAvac QIAvac 6S QIAquick QIAvac Luer Adapter Set 3c. QIAquick 4. DNA QIAquick 1 800 µl 800 µl 5. 750 µl Buffer PE QIAquick 6. QIAquick 2 ml 17,900 x g 13,000 rpm 1 Buffer PE 7. 1.5 mlqiaquick 8. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 7.0 8.5 QIAquick 17,900 x g 13,000 rpm 1DNA 30 µl QIAquick 1 DNA QIAquick 50 µl 48 µl 30 µl 28 µl ph ph 7.0 8.5 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 6 QIAquick Spin 03/2008

9. DNALoading Dye 5 DNA Loading dye 3 bromophenol blue xylene cyanol orange GDNA Table 2 Handbook 15 QIAquick Spin 03/2008 7

QIAquick Nucleotide Removal Kit Handbook 8 DIG DNA 17 10 96 100 Buffer PE 15 25 1. 10Buffer PN 50 µl 500 µl Buffer PN 100 bp DNA 5 Buffer PN 2. 2 mlqiaquick 3. DNA QIAquick 6,000 rpm 1 4. QIAquick 2 ml QIAquick 5. QIAquick 500 µl Buffer PE 6,000 rpm 1 500 µl Buffer PE QIAquick 750 µl Buffer PE 6,000 rpm 1 8 QIAquick Spin 03/2008

6. QIAquick 17,900 x g 13,000 rpm 1 Buffer PE 7. 1.5 mlqiaquick 8. DNA 100 200 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 7.0 8.5 QIAquick 17,900 x g 13,000 rpm 1DNA 30 50 µl QIAquick 1 DNA QIAquick ph ph 7.0 8.5 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 9. DNALoading Dye 5 DNA Loading dye 3 bromophenol blue xylene cyanol orange GDNA Table 2 Handbook 15 QIAquick Spin 03/2008 9

QIAquick Gel Extraction Kit TAE TBE 70 bp 10 kb DNA 1 400 mg DNA Handbook 8 DNA 3 Buffer QG 6 MinElute Reaction Cleanup Kit Buffer QG ph 7.5 96 100 Buffer PE 17,900 x g 13,000 rpm 15 25 1. DNA 2. 100 mg 100 µl 3Buffer QG 100 mg 300 µl Buffer QG 2 6Buffer QG 1 QIAquick 400 mg 400 mg 2 QIAquick 3. 50 10 2 3 2 10 QIAquick Spin 03/2008

4. Buffer QG 3M ph 5.0 10 µl DNA QIAquick ph 7.5 Buffer QG ph ph 7.5 ph ph DNA ph 5. 100 mg 100 µl 500 bp 4 kb DNA 500 bp 4 kb DNA 6. 2 mlqiaquick 7. DNA QIAquick 1 1 800 µl 800 µl 8. QIAquick 9. 500 µl Buffer QG QIAquick1 DNA in vitro 10. 750 µl Buffer PE QIAquick1 DNA Buffer PE 2 5 11. QIAquick 17,900 x g 13,000 rpm 1 Buffer PE 12. QIAquic 1.5 ml QIAquick Spin 03/2008 11

13. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 7.0 8.5 QIAquick 1 DNA 30 µl QIAquick 1 1 DNA QIAquick 50µl 48 µl 30 µl 28 µl ph ph 7.0 8.5 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 14. DNALoading Dye 5 DNA Loading dye 3 bromophenol blue xylene cyanol orange GDNA Table 2 Handbook 15 12 QIAquick Spin 03/2008

QIAquick Gel Extraction Kit QIAquick QIAvac Luer Adaptors QIAvac 6S/QIAvac 24 Plus TAETBE 70 bp 10 kb DNA 1 400 mg DNA Handbook 8 DNA 3 Buffer QG 4 7 MinElute Reaction Cleanup Kit Buffer QG ph 7.5 96 100 Buffer PE 1. DNA 2. 100 mg 100 µl 3Buffer QG 100 mg 300 µl Buffer QG 2 6 Buffer QG 1 QIAquick 400 mg 400 mg 2 QIAquick 3. 50 10 2 3 2 4. QIAquick 4a 4b 4c QIAquick Spin 03/2008 13

4a. QIAvac 24 PlusHandbook 33Figure 7 QIAvac 24 Plus 24 QIAquick QIAvac 24 Plus 4b. QIAvac 6S Handbook 34Figure 8 QIAvac 6S QIAvac QIAvac Luer Adapter QIAvac 6S QIAvac QIAvac QIAvac 6S QIAquick QIAvac Luer Adapter Set 4c. QIAquick 5. Buffer QG 3M ph 5.0 10 µl DNA QIAquick ph 7.5 Buffer QG ph ph 7.5 ph DNA ph 6. 100 mg 100 µl 500 bp 4 kb DNA 500 bp 4 kb DNA 7. DNA QIAquick 1 800 µl 800 µl 8. 500 µl Buffer QG QIAquick DNA in vitro 14 QIAquick Spin 03/2008

9. 0.75 ml Buffer PE QIAquick DNA Buffer PE 2 5 10. QIAquick 1.5 ml 2 ml 17,900 x g 13,000 rpm 1 Buffer PE 11. 1.5 mlqiaquick 12. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 7 8.5 QIAquick 17,900 x g 13,000 rpm 1 DNA 30 µl QIAquick 1 1 DNA QIAquick 50 µl 48 µl 30 µl 28 µl ph ph 7.0 8.5 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 13. DNALoading Dye 5 DNA Loading dye 3 bromophenol blue xylene cyanol orange GDNA Table 2 Handbook 15 QIAquick Spin 03/2008 15

a Buffer PE b c d e ph f >400 mg Buffer PE Buffer PE DNA Buffer EB 10 mm Tris Cl ph 8.5 Elution in low-salt solutions Handbook 13 QIAquick 30 µl Buffer QG50 2 3 DNA ph Buffer QG DNA 10 µl 3M ph 5.0 DNA ph 10 µl QIAquick 400 mg 70 80 400 mg QIAquick QIAquick Gel Extraction Kit PCR QIAquick PCR Purification Kit 16 QIAquick Spin 03/2008

PCR g PCR PCR/Gel h i DNA a b c PCR 10 DNA QIAquick 0.5 ml Buffer QG 15 25 1 ph Buffer QG PB 20 µl 3M ph 5.0 DNA ph 10 µl 750 µl Buffer PE 5 QIAquick QIAquick 17,900 x g 13,000 rpm 1 400 mg Buffer QG QIAquick Spin 03/2008 17

PCR d e ssdna 20 bp QIAquick 750 µl 35 35 g/100 ml Buffer PE DNA ssdna 95 2 15 25 10 mm NaCl 10 mm Tris DNA DNA 18 QIAquick Spin 03/2008

Memo QIAquick Spin 03/2008 19

Trademarks: QIAGEN, QIAquick, MinElute (QIAGEN Group). QIAGEN 2007 2008 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 2301417 06/2008 Sample & Assay Technologies