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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1 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

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

3 QIAquick PCR Purification Kit PCR Handbook 8 DNA PCR MinElute Reaction Cleanup Kit QIAquick 100 bp 10 kb Buffer PE 17,900 x g 13,000 rpm ph indicator I Buffer PB 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 µl 3. 2 mlqiaquick 4. DNA QIAquick QIAquick QIAquick Spin 03/2008 3

4 µl Buffer PE QIAquick QIAquick 1 Buffer PE mlqiaquick 9. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph QIAquick 1 DNA 30 µl QIAquick 1 1 DNA QIAquick 50 µl 48 µl 30 µl 28 µl ph ph 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

5 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 Buffer PE ph indicator I Buffer PB 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 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

6 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 µl 800 µl µl Buffer PE QIAquick 6. QIAquick 2 ml 17,900 x g 13,000 rpm 1 Buffer PE mlqiaquick 8. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 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 ph DNA 20 DNA TE 10 mm Tris Cl 1 mm EDTA ph 8.0 EDTA 6 QIAquick Spin 03/2008

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

8 QIAquick Nucleotide Removal Kit Handbook 8 DIG DNA Buffer PE Buffer 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 µl Buffer PE QIAquick 750 µl Buffer PE 6,000 rpm 1 8 QIAquick Spin 03/2008

9 6. QIAquick 17,900 x g 13,000 rpm 1 Buffer PE mlqiaquick 8. DNA µl Buffer EB 10 mm Tris Cl ph 8.5 ph QIAquick 17,900 x g 13,000 rpm 1DNA µl QIAquick 1 DNA QIAquick ph ph 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

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

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

12 13. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph QIAquick 1 DNA 30 µl QIAquick 1 1 DNA QIAquick 50µl 48 µl 30 µl 28 µl ph ph 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 QIAquick Spin 03/2008

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

14 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 µl DNA QIAquick ph 7.5 Buffer QG ph ph 7.5 ph DNA ph mg 100 µl 500 bp 4 kb DNA 500 bp 4 kb DNA 7. DNA QIAquick µl 800 µl µl Buffer QG QIAquick DNA in vitro 14 QIAquick Spin 03/2008

15 ml Buffer PE QIAquick DNA Buffer PE QIAquick 1.5 ml 2 ml 17,900 x g 13,000 rpm 1 Buffer PE mlqiaquick 12. DNA 50 µl Buffer EB 10 mm Tris Cl ph 8.5 ph 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 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/

16 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 QG DNA ph Buffer QG DNA 10 µl 3M ph 5.0 DNA ph 10 µl QIAquick 400 mg mg QIAquick QIAquick Gel Extraction Kit PCR QIAquick PCR Purification Kit 16 QIAquick Spin 03/2008

17 PCR g PCR PCR/Gel h i DNA a b c PCR 10 DNA QIAquick 0.5 ml Buffer QG 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 mg Buffer QG QIAquick Spin 03/

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

19 Memo QIAquick Spin 03/

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

HotStarTaq Plus PCRプロトコールとトラブルシューティング( /2008) February 2008 HotStarTaq Plus PCR HotStarTaq Plus DNA Polymerase HotStarTaq Plus Master Mix Kit PCR HotStarTaq Plus DNA Polymerase PCR 2 HotStarTaq Plus DNA Polymerase Q-Solution PCR 6 HotStarTaq Plus