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 15 18 Sample & Assay Technologies
HotStarTaq Plus DNA Polymerase PCR HotStarTaq Plus DNA Polymerase 95 5 6 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. 201900 1. 10x CoralLoad PCR Buffer10x PCR Buffer dntp 25 mm MgCl 2 2. 1 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
1. HotStarTaq Plus DNA Polymerase 10x CoralLoad PCR Buffer* 10 µl 1x 10x PCR Buffer* 25 mm MgCl 2 2 2 dntp 10 mm 2 µl 200 µm dntp A 0.1 0.5 µm B 0.1 0.5 µm HotStarTaq Plus DNA 0.5 µl 2.5 Polymerase RNase DNA DNA 1 µg 4 100 µl * 15 mm MgCl 2 Handbook 36 Table 16 2. Mg 2+ Mg 2+ mm 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 25 mm MgCl 2 µl 0 2 4 6 8 10 12 14 Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 1.5 mm HotStarTaq Plus PCR 02/2008 3
3. PCR PCR HotStarTaq Plus DNA Polymerase DNA 4. DNA 1 µg RT-PCR PCR 10 Handbook 40 Appendix E 5. 6 100 µl 6. 95 5 PCR 5 3. PCR 5 95 HotStarTaq Plus DNA Polymerase 3 0.5 1 94 0.5 1 50 68 T m 5 Handbook 36 Appendix B 1 72 1 kb PCR 1 kb DNA 1 25 35 Handbook 39 Appendix C 10 72 4 HotStarTaq Plus PCR 02/2008
7. PCR 2 8 20 8. CoralLoad PCR Buffer PCR CoralLoad PCR Buffer 4 4. %TAE TBE 0.8 500 bp 270 bp 80 bp <10 bp 1.0 300 bp 220 bp 40 bp <10 bp 1.5 250 bp 120 bp 20 bp <10 bp 2.0 100 bp 110 bp <10 bp <10 bp 3.0 50 bp 100 bp <10 bp <10 bp HotStarTaq Plus PCR 02/2008 5
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 + + 2 kb 0.32 kb C M + + D M + + 1.5 kb 0.5 kb Q-Solution + Q-Solution M 6 HotStarTaq Plus PCR 02/2008
HotStarTaq Plus DNA Polymerase 95 5 6 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. 201900 1. 10x CoralLoad PCR Buffer10x PCR Buffer dntp Q-Solution Q-Solution MgCl 2 2. 8 5 HotStarTaq Plus DNA Polymerase DNA PCR PCR 10 DNA HotStarTaq Plus PCR 02/2008 7
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 0.1 0.5 µm B 0.1 0.5 µm HotStarTaq Plus 0.5 µl 2.5 DNA Polymerase RNase DNA DNA 1 µg 4 100 µl * 15 mm MgCl 2 Handbook 36 Table 16 3. 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
5. 100 µl 6. 95 5 PCR 5 PCR 6 6. PCR 5 95 HotStarTaq Plus DNA Polymerase 3 0.5 1 94 0.5 1 50 68 T m 5 Handbook 36 Appendix B 1 72 1 kb PCR 1 kb DNA 1 25 35 Handbook 39 Appendix C 10 72 7. PCR 2 8 20 HotStarTaq Plus PCR 02/2008 9
8. CoralLoad PCR Buffer PCR CoralLoad PCR Buffer 7 7. %TAE TBE 0.8 500 bp 270 bp 80 bp <10 bp 1.0 300 bp 220 bp 40 bp <10 bp 1.5 250 bp 120 bp 20 bp <10 bp 2.0 100 bp 110 bp <10 bp <10 bp 3.0 50 bp 100 bp <10 bp <10 bp 10 HotStarTaq Plus PCR 02/2008
HotStarTaq Plus DNA Polymerase PCR DNA HotStarTaq Plus DNA Polymerase 95 5 5 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. 201900 HotStarTaq Plus PCR 02/2008 11
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 2+ 8. HotStarTaq Plus DNA Polymerase 10x CoralLoad PCR Buffer* 5 µl 1x 10x PCR Buffer* 25 mm MgCl 2 9 9 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 16 12 HotStarTaq Plus PCR 02/2008
9. Mg 2+ Mg 2+ mm 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 25 mm MgCl 2 µl 0 2 4 6 8 10 12 14 Mg 2+ 1x PCR Buffer 1x CoralLoad PCR Buffer 1.5 mm 3. 30 µl PCR 4. 5 50 µl 5. 95 5 PCR 5 10. PCR 5 95 HotStarTaq Plus DNA Polymerase 3 0.5 1 94 0.5 1 50 68 T m 5 Handbook 36 Appendix B 1 72 1 kb PCR 1 kb DNA 1 45 50 Handbook 39 Appendix C 10 72 HotStarTaq Plus PCR 02/2008 13
6. PCR 2 8 20 7. CoralLoad PCR Buffer PCR CoralLoad PCR Buffer 11 11. %TAE TBE 0.8 500 bp 270 bp 80 bp <10 bp 1.0 300 bp 220 bp 40 bp <10 bp 1.5 250 bp 120 bp 20 bp <10 bp 2.0 100 bp 110 bp <10 bp <10 bp 3.0 50 bp 100 bp <10 bp <10 bp 14 HotStarTaq Plus PCR 02/2008
HotStarTaq Plus Master MixPCR HotStarTaq Plus Master Mix 95 5 6 HotStarTaq Plus Master Mix MgCl2 1.5 mm Mg 2+ 25 mm MgCl 2 PCR QIAquick PCR Purification Kit MinElute PCR Purification Kit CoralLoad Concentrate DNA PCR 1. 2. 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 02/2008 15
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 0.1 0.5 µm B 0.1 0.5 µm CoralLoad 2 µl 1x CoralLoad Concentrate 10x Concentrate RNase DNA DNA 20 µl 4 200 ng 20 µl * 1.5 mm MgCl 2 5. 6 50 µl 6. 95 5 PCR 5 13 17 16 HotStarTaq Plus PCR 02/2008
13. PCR 5 95 HotStarTaq Plus DNA Polymerase 3 0.5 1 94 0.5 1 50 68 T m 5 Handbook 36 Appendix B 1 72 1 kb PCR 1 kb DNA 1 25 35 Handbook 39 Appendix C 10 72 7. PCR 2 8 20 8. CoralLoad Concentrate PCR CoralLoad PCR Buffer 14 14. %TAE TBE 0.8 500 bp 270 bp 80 bp <10 bp 1.0 300 bp 220 bp 40 bp <10 bp 1.5 250 bp 120 bp 20 bp <10 bp 2.0 100 bp 110 bp <10 bp <10 bp 3.0 50 bp 100 bp <10 bp <10 bp HotStarTaq Plus PCR 02/2008 17
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 6 0.1 0.5 µm 0.1 µm PCR PCR * Handbook 35 Appendix A PCR PCR nested PCR 2 PCR Handbook 39 Appendix D 25 mm MgCl 2 0.5 mm 1.5 5.0 mm Mg 2+ PCR 3 2 100 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
i j k l m n RT o DNA PCR p PCR q 2 30 60 PCR Handbook 42 Appendix F 94 30 60 PCR 6 4 9 5 13 95 5 1 DNA PCR o DNA PCR Nested PCR 2 PCR Handbook 39 Appendix D Handbook 36 Appendix B RT-PCR 10 30 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 02/2008 19
a HotStarTaq Plus DNA Polymerase b PCR c d e a b 95 5 Q-Solution PCR 6 2 30 60 PCR Handbook 42 Appendix F 0.1 0.5 µm 0.1 µm PCR PCR * Handbook 36 Appendix B Handbook 35 Appendix A PCR PCR PCR 10 3 10 4 1 µl Nested PCR Handbook 39 Appendix D PCR DNA PCR DNA PCR * MSDS material safety data sheet 20 HotStarTaq Plus PCR 02/2008
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 95 5 3 25 mm MgCl 2 0.5 mm 1.5 5.0 mm MgCl 2 PCR 3 2 0.1 0.5 µm 0.1 µm PCR PCR Handbook 36 Appendix B HotStarTaq Plus PCR 02/2008 21
Memo 22 HotStarTaq Plus PCR 02/2008
Memo HotStarTaq Plus PCR 02/2008 23
Trademarks: QIAGEN, QIAquick, CoralLoad, HotStarTaq, MinElute, ProofStart, Q-Solution (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 2301467 10/2008 Sample & Assay Technologies