HotStarTaq Plus PCRプロトコールとトラブルシューティング( /2008)
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1 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 DNA Polymerase PCR 11 HotStarTaq Plus Master Mix PCR Sample & Assay Technologies
2 HotStarTaq Plus DNA Polymerase PCR HotStarTaq Plus DNA Polymerase DNA PCR PCR QIAquick PCR Purification Kit MinElute PCR Purification Kit CoralLoad PCR Buffer dntp 10 mm dntp 20 QIAGEN PCR dntp 10 mm Cat. no x CoralLoad PCR Buffer10x PCR Buffer dntp 25 mm MgCl HotStarTaq Plus DNA Polymerase DNA PCR PCR 10 DNA Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 2 Mg 2+ 2 HotStarTaq Plus PCR 02/2008
3 1. HotStarTaq Plus DNA Polymerase 10x CoralLoad PCR Buffer* 10 µl 1x 10x PCR Buffer* 25 mm MgCl dntp 10 mm 2 µl 200 µm dntp A µm B µm HotStarTaq Plus DNA 0.5 µl 2.5 Polymerase RNase DNA DNA 1 µg µl * 15 mm MgCl 2 Handbook 36 Table Mg 2+ Mg 2+ mm mm MgCl 2 µl Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 1.5 mm HotStarTaq Plus PCR 02/2008 3
4 3. PCR PCR HotStarTaq Plus DNA Polymerase DNA 4. DNA 1 µg RT-PCR PCR 10 Handbook 40 Appendix E µl PCR 5 3. PCR 5 95 HotStarTaq Plus DNA Polymerase T m 5 Handbook 36 Appendix B kb PCR 1 kb DNA Handbook 39 Appendix C HotStarTaq Plus PCR 02/2008
5 7. PCR CoralLoad PCR Buffer PCR CoralLoad PCR Buffer 4 4. %TAE TBE bp 270 bp 80 bp <10 bp bp 220 bp 40 bp <10 bp bp 120 bp 20 bp <10 bp bp 110 bp <10 bp <10 bp bp 100 bp <10 bp <10 bp HotStarTaq Plus PCR 02/2008 5
6 HotStarTaq Plus DNA Polymerase Q-Solution PCR PCR Q-Solution Q-Solution DNA PCR Q-Solution Q-Solution DMSO PCR Q-Solution PCR A Q-Solution B Q-Solution PCR C Q-Solution PCR D Q-Solution Q-Solution Q-Solution Q-Solution A M + + B M kb 0.32 kb C M + + D M kb 0.5 kb Q-Solution + Q-Solution M 6 HotStarTaq Plus PCR 02/2008
7 HotStarTaq Plus DNA Polymerase Q-Solution Q-Solution DNA PCR PCR QIAquick PCR Purification Kit MinElute PCR Purification Kit CoralLoad PCR Buffer dntp 10 mm dntp 20 QIAGEN PCR dntp 10 mm Cat. no x CoralLoad PCR Buffer10x PCR Buffer dntp Q-Solution Q-Solution MgCl HotStarTaq Plus DNA Polymerase DNA PCR PCR 10 DNA HotStarTaq Plus PCR 02/2008 7
8 5. HotStarTaq Plus DNA Polymerase Q-Solution 10x CoralLoad PCR Buffer* 10 µl 1x 10x PCR Buffer* 5x Q-Solution 20 µl 1x dntp 2 µl 200 µm dntp 10 mm A µm B µm HotStarTaq Plus 0.5 µl 2.5 DNA Polymerase RNase DNA DNA 1 µg µl * 15 mm MgCl 2 Handbook 36 Table PCR PCR HotStarTaq Plus DNA Polymerase DNA 4. DNA 1 µg RT-PCR PCR 10 Handbook 40 Appendix E 8 HotStarTaq Plus PCR 02/2008
9 µl PCR 5 PCR 6 6. PCR 5 95 HotStarTaq Plus DNA Polymerase T m 5 Handbook 36 Appendix B kb PCR 1 kb DNA Handbook 39 Appendix C PCR HotStarTaq Plus PCR 02/2008 9
10 8. CoralLoad PCR Buffer PCR CoralLoad PCR Buffer 7 7. %TAE TBE bp 270 bp 80 bp <10 bp bp 220 bp 40 bp <10 bp bp 120 bp 20 bp <10 bp bp 110 bp <10 bp <10 bp bp 100 bp <10 bp <10 bp 10 HotStarTaq Plus PCR 02/2008
11 HotStarTaq Plus DNA Polymerase PCR DNA HotStarTaq Plus DNA Polymerase E. coli 5S rrna HPLC DNA PCR PCR QIAquick PCR Purification Kit MinElute PCR Purification Kit CoralLoad PCR Buffer DNA 20 µl 1x PCR Buffer PCR PCR 80 dntp 10 mm dntp 20 QIAGEN PCR dntp 10 mm Cat. no HotStarTaq Plus PCR 02/
12 1. 10x CoralLoad PCR Buffer10x PCR Buffer dntp 25 mm MgCl 2 PCR 2. 8 DNA PCR PCR 10 DNA Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 9 Mg HotStarTaq Plus DNA Polymerase 10x CoralLoad PCR Buffer* 5 µl 1x 10x PCR Buffer* 25 mm MgCl dntp 1 µl 200 µm dntp 10 mm A 0.2 µm B 0.2 µm RNA 50 ng HotStarTaq Plus 1 µl 5 DNA Polymerase RNase 1 x PCR Buffer 20 µl 50 µl * 15 mm MgCl 2 Handbook 36 Table HotStarTaq Plus PCR 02/2008
13 9. Mg 2+ Mg 2+ mm mm MgCl 2 µl Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 1.5 mm µl PCR µl PCR PCR 5 95 HotStarTaq Plus DNA Polymerase T m 5 Handbook 36 Appendix B kb PCR 1 kb DNA Handbook 39 Appendix C HotStarTaq Plus PCR 02/
14 6. PCR CoralLoad PCR Buffer PCR CoralLoad PCR Buffer %TAE TBE bp 270 bp 80 bp <10 bp bp 220 bp 40 bp <10 bp bp 120 bp 20 bp <10 bp bp 110 bp <10 bp <10 bp bp 100 bp <10 bp <10 bp 14 HotStarTaq Plus PCR 02/2008
15 HotStarTaq Plus Master MixPCR HotStarTaq Plus Master Mix HotStarTaq Plus Master Mix MgCl2 1.5 mm Mg mm MgCl 2 PCR QIAquick PCR Purification Kit MinElute PCR Purification Kit CoralLoad Concentrate DNA PCR HotStarTaq Plus Master Mix12 PCR 10 µl HotStarTaq Plus Master Mix HotStarTaq Plus DNA Polymerase 3. Master MixPCR 4. PCR DNA 20 µl 200 ng RT-PCR PCR 10 Handbook 40 Appendix E HotStarTaq Plus PCR 02/
16 12. HotStarTaq Plus Master Mix HotStarTaq Plus Master Mix 2 x 10 µl 1 unit HotStarTaq Plus DNA Polymerase 1 x PCR Buffer* 200 µm dntp A µm B µm CoralLoad 2 µl 1x CoralLoad Concentrate 10x Concentrate RNase DNA DNA 20 µl ng 20 µl * 1.5 mm MgCl µl PCR HotStarTaq Plus PCR 02/2008
17 13. PCR 5 95 HotStarTaq Plus DNA Polymerase T m 5 Handbook 36 Appendix B kb PCR 1 kb DNA Handbook 39 Appendix C PCR CoralLoad Concentrate PCR CoralLoad PCR Buffer %TAE TBE bp 270 bp 80 bp <10 bp bp 220 bp 40 bp <10 bp bp 120 bp 20 bp <10 bp bp 110 bp <10 bp <10 bp bp 100 bp <10 bp <10 bp HotStarTaq Plus PCR 02/
18 a HotStarTaq Plus DNA Polymerase b c PCR d e f Mg 2+ g h 95 5 PCR 15 PCR dntps HotstarTaq Plus Master Mix HotStarTaq Plus Master Mix 1 1 Q-Solution PCR µm 0.1 µm PCR PCR * Handbook 35 Appendix A PCR PCR nested PCR 2 PCR Handbook 39 Appendix D 25 mm MgCl mm mm Mg 2+ PCR µl 2.5 HotStarTaq Plus DNA Polymerase 0.5 HotStarTaq Plus DNA Polymerase HotStarTaq Plus Master Mix 20 µl 10 µl 5 Handbook 39 Appendix C * MSDS material safety data sheet 18 HotStarTaq Plus PCR 02/2008
19 i j k l m n RT o DNA PCR p PCR q PCR Handbook 42 Appendix F PCR DNA PCR o DNA PCR Nested PCR 2 PCR Handbook 39 Appendix D Handbook 36 Appendix B RT-PCR PCR 10 Handbook 40 Appendix E DNA 4 kb DNA Handbook 35Appendix AQIAGEN Taq DNA Polymerase HotStar HiFidelity Polymerase PCR Taq PCR HandbookHotStar HiFidelity PCR HandbookQIAGEN Long Range PCR Kit PCR PCR HotStarTaq Plus PCR 02/
20 a HotStarTaq Plus DNA Polymerase b PCR c d e a b 95 5 Q-Solution PCR PCR Handbook 42 Appendix F µm 0.1 µm PCR PCR * Handbook 36 Appendix B Handbook 35 Appendix A PCR PCR PCR µl Nested PCR Handbook 39 Appendix D PCR DNA PCR DNA PCR * MSDS material safety data sheet 20 HotStarTaq Plus PCR 02/2008
21 c d HotStarTaq Plus DNA Polymerase e f Mg 2+ g h 100 µl 2.5 HotStarTaq Plus DNA Polymerase HotStarTaq Plus Master Mix 20 µl 10 µl mm MgCl mm mm MgCl 2 PCR µm 0.1 µm PCR PCR Handbook 36 Appendix B HotStarTaq Plus PCR 02/
22 Memo 22 HotStarTaq Plus PCR 02/2008
23 Memo HotStarTaq Plus PCR 02/
24 Trademarks: QIAGEN, QIAquick, CoralLoad, HotStarTaq, MinElute, ProofStart, Q-Solution (QIAGEN Group). QIAGEN QIAGEN, all rights reserved Forefront Tower II Tel: Fax: /2008 Sample & Assay Technologies
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製品コード 6126/6127 BKL Kit (Blunting Kination Ligation Kit) 説明書 PCR 産物を平滑末端ベクターにクローニングする場合 使用するポリメラーゼ酵素の種類により 3' 末端に余分に付加された塩基を除去し さらに 5' 末端をリン酸化する必要があります 本製品は これらの一連の反応を簡便に短時間に行うためのキットです PCR 産物の末端平滑化とリン酸化を同時に行うことにより
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DynalTransplantDiagnostic.indd
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(1) / 130 g 25 g 520% 170 g 30 g 560% 70 mg 600 mg 11.6% 0 10.5 mg 0% (1) (2) / 50100 g 25 g 200400% 50100 g 30 g 167333% 5001000 mg 600 mg 83167% 1020 mg 10.5 mg 95190% (2) / (1) 45.6 g 30 g 152% (2)
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