PCR 実験の手引き
Contents
1. CO G 3 PCR PCR2. PCR 2 12 lahaha PCR EL LL LL
3. 1 PCR GenBank DDB NCBI
4. PCR PCR PCR PCR 1 DNA 0 PCR 1 lahaha 8 0 2ml 8 Vortex 20 200 1000
5. 70C 0 5g g mL EF IafRa PCR ETL LLL 5 mm
6. PCR 1 3 5 20 TakaRa Pamofe PCR 1 w 1 1 5mL 11 PCR
7. nn mm mn 1 Ty Rm NT 5 3 E 1XxTAE DNA DNA
8. NCBI NCBI web page NCBI http www ncbi nlm nih gqov a NC Search All Databas 4 NCBI Home Welcome to NCBI Popular Resources A OQT Iniormabhon adrances saence and LAS The 3 al Center for Bio ology inf t d d BLAST aith by provdin cess to biomedical and genomic information Bookshe sene amp Biloassa bout the NCBI Mission Organization Research RSS Feeds Genome Data amp s vare hucieotide F OlAIN NA amp RNA 0 Get Started Protein Domains amp Structures PubCher Ge Tools Analyze data using NCBI softw PubMed _ Downlosds MM data or soWw PublMed How To s Lesrm how 0 accomplish specific s at NCBI SNP Submissions Submit data to Ge eave Of po ete NCBI News 14 ffaR8 2
9. 9 4 A T 6G C DNA Deoxyribonucleic acid A G T O 4 p k3 DNA DNA 1 5 DNA DNA DNA DNA 3 DNA
10. CU 2 OCOD CO
11. DNA Hot Start 20 TakaRa Pramome 5 1 PCR DNA DNA DNA DNA PCR DNA DNA DNA
12. PCR 9 6 wy MM ul DNA 3 Agarose LO03 5003 SeaKem LE Agarose F0004 NuSieve 3 1 Agarose F0090 Reliant Gel System DNA
13. DNA PCR DNase RNase PCR DNase RNase 8 0 2 ml 8 strip tube Individual Flat Caps NJ600 TakaRa Pramome 1 5 ml 0 2 ml 8 8 1 5 ml 0 2 ml PCR 1 5 ml 0 2 ml
14. 25 40 DNA dQNTP DNA 2 PCR DNA DNA p 2 DNA pH Mg DNA buffer
15. DNA 8 DNA TAE buffer 2 PCR PCR PCR UV DNA ey UV
16. 16 TakaRa Pramome 3 DNase EDTA DNase TE buffer TE buffer 10mM Tris HC1 pH8 0 0 1mM EDTA DNase EDTA DNase 2 PCR
17. dNTP DNA DNA DNA DNA DNA DNA RNA PCR PCR DNA DNA DNA PCR DNA DNA DNA PCR 1 DNA DNA DNA 2 DNA DNA
18. dNTP deoxyribonucleotide 5 triphosphate DNA dATP dGTP dCTP dTTP 4 Mg TalaRa maeofe 4 1 DNA 1 DNA DNA DNA DNA DNA 5
19. 95 C 1 3 1 lt 230 lahaha PCR 1 Tm 5C Tm Tm DNA 50 DNA DNA 1kbp 1
20. DNA 2 PCR Ss aa 06 DNA PCR DNA PCR
21. DNA 3 5 exonuclease Pol 1 5 i GATCTG CTAGATCGGAT 5 DNA 3 5 exonuclease G EE GATcT _ CTAGATCGGAT 5 DNA GATCT SS CTAGATCGGAT 5 DNA A GATCTA CTAGATCGGAT 5 DNA ma 2 TakaRa mweofe 3 Hot Start PCR Hob Sbart PCRIG DNA DNA DNA PCR PCR
22. 2013 4 1 TaKaRa TEL 07 543 6116 FAX 07 543 197 TEL 03 3271 6553 FAX 03 32 71 262 TEL 07 565 6969 FAX 07 565 6995 1304
23. 57 9 CO 3 5 68 C 72 C DNA DNA 57 CY DNA 5 3 3 5 3 n 2 n DNA 100 57 cy 3 4 n 2n DNA 100 3 PCR 1 PCR DNA DNA DNA DNA DNA
24. TakaRa Pramome 3 FlashGel System for DNA ha DNA 1kb 1kb EE _ Da o DNA PCR Size Range bp SeaKem LE LO3 H14 Size Range DD NuSieve 3 1 1 000 23 000 0 6 w v 500 1 000 3 0 w v 800 10 000 0 8 w v 100 500 4 0 w v 400 8 000 1
25. DNA PCR DNA DNA PCR 6 A 13 lahaha PCR LL LL gt 1 PCR 1 2 3
26. PCR 1 11 33 5L 368 5L 10X PCR 5 0L 55 0L dNTP Mixture 2 5mW 4 0L 44 0L F Primer 1 0L 11 0L R Primer 1 0L 11 0L PCR 0 5L 5 5uL Total 45 0L 495L PCR PCR 0 2nL PCR PCR 0 2mL 45ppL 5zL 3 3
27. __ A Hind III digest pHY Marker lahaha PCR HHHOUL 4 PCR 1 PCR PCR PCR M 1 3 4 1 PCR 2 PCR 3 PCR 4 PCR COD 2
28. 57 3 9 57 94C 98 DNA 57 1 2 DNA 50 65C DNA DNA DNA DNA A T G C 5 3 3 5 3 3 50C 65 A T G C 5 9 DNA 3 5 DNA DNA 5 Takaha maeofe 3 DNA DNA DNA DNA
29. 1 DNA PCR 0 MA 00 2 Uracil N glycosylase UNG TaKaRa PCR Carryover Prevention Kit code 6088 PCR PCR DNA 15 1 1 1 1 UNG 25 10 95 3 dU plus dNTP Mixture PCR PCR PCR dTTP dUTP
30. PCR IafRa PCR EL LLL LLL Tm nearest neighbor Tm Tm Tm C 60 8 0 41X GC 500 n Tm CC 4X G C 2 Xx A T 35 2n GC GC n A G T C IMC Web Page http www takara biio co jp TakaRa 9
31. bp ER a LO3 Hi14 SeaKem LE NuSieve 3 1 196 1 596 296 396 496 696 lt 6 100 2 800 1 300 650 350 120 BPB LO3 Hi14 SeaKem LE NuSieve 3 1 ss lt 196 1 5 2 3 4 6 500 300 150 80 40 20 1xTAE buffer DNA bp bp 100 1000 10000 100000 1000 1 11 1L 1 AL L I 1000 20bp DNA Ladder 700 a _ 750 50 bp DNA Ladder 100 bp DNA Ladder ee Ee 0 200 bp DNA Ladder 0 250 bp DNA Ladder 500 bp DNA Ladder a 1kbDNA Ladder 25kbDNA Ladder ioo 100 Wide Range DNA Ladder 100bp DNA Ladder Wide Range DNA Ladder 50 10 000 bp es 100 2 000 bp 2 100 2 000 bp bp bp 100 5 000 bp 23130 5 s 9416 X174 Hinc lldigest 2016 X174 Hae digest 8 1360 1107 pHY Marker _ 0 202 A Hind digest BstP digest EcoT14 digest A EcoT14 Bgl ll digest 489 267 Dye Ready to use
32. lahaha PCR 1 2 2 1 3 2 3 4 4 1 3 4 gt 3
33. PCR DNA mRNA miRNA PCR DNA gt Rs lt gt DNA 6 PCR lt gt gt 6 8 PCR 6 qPCR 6 6 mRNA miRNA lahaha PCR gt 1 DNA DNA DNA Deoxyribonucleic acid DNA DNA DNA
34. 5zL 4 PCR PCR PCR PCR PCR IafRa PCR 3 LL LI LL DNA UV DNA 1 SS 2 2 1 XTAE buffer Tris Acetate EDTA buffer 20 50 lt gt Tris hydroxymethyl aminomethane 242g 57 1 ml 0 5M EDTA 100mL 1L 50 50 1XTAE 10mg m1 Ethidium
35. A T G C dNTP 1 DNA DNA DNA 5 1 DNA PCR DNA DNA DNA DNA DNA PCR DNA DNA DNA 2 DNA DNA 3 RNA 97 4 DNA 5 RNA DNA DNA DNA
36. PCR 9 UNG PCR DNA 28 TakaRa mweofe 6 lahaha PCR PCR A4 TaKaRa PCR Thermal Cycler Dice 7Touch PCR TP350 580 000 96 TaKaRa PCR Ihermal Cycler Dice Gradient PCR TP600 TaKaRa PCR Ihermal Cycler Dice Standard IE PR a 24
37. 24 TakaRa Pramome UV DNA 1XTAE 0 5g mL 1 TAE SYBR Green 1 15 UV DNA PCR BPB XC
38. a TaKaRa PCR Thermal Cycler Dice mini PCR TP100 480 000 PCR 8 3 0 2 ml 8 strip tube individual Flat Caps NJ600 18 000 ee 0 2 ml Hi Tube Dome Cap 2 NJ200 1 000 Y8 500 FF 0 2 ml Hi Tube Flat Cap 9 NJ202 1 000 8 500 0 2 ml Hi 8 Tube 8 NJ300 1 15 000 strips 0 2 ml Hi 8 Dome Cap 5 NJ301 4 000 strips 5 J 0 2 ml Hi 8 Flat Cap nso 4 000 50 8 iaKeRa POR Hi Gaps 8 Mupid 2plus 40 762 Agarose L03 TAKARAI MG 5003B 78 000 100 bp 23 000 bp 50001 11 300 DNA 50000 125g 32 000 SeaKem LE Agarose 125g 50004 92 000 50091 22 900 leve 3 yi Te ET 62 700_ NuSieve 3 1 Agarose 50090 62 700 50094 223 000 PCR TaKaRa PCR Carryover P
39. i fd ns 4 lJ ae mL http www insilicobioloqy cO 2 Primer 3 http frodo wi mit edu DNA PCR NCBI Primer BLAST http www ncbi nlm nih gov tools primer blast index cgi LINK_LOC BlastHome Primer3 BLAST Primer BLAST BLAST Basic Local Alignment Search Tool DNA
40. 5 3 DNA DNA DNA 5 3 5 CH2 DNA DNA QO 57 lt DNA 3 DNA 3 57 5 HO P O JHTLLTLILTTLIITIITII 3 sy CH gt DNA O QO 4 PCR Lb EE NN bt LL ffaR8 DNA PCR DNA DNA DNA DNA DNA a ee RNA DNA RE DNA
41. 7 IafRa PCR 1 HHHOUL PCR PCR PCR X R PCR X X X 1 a PCR i My PCR
42. DNA 3 gt RNA DONA 4 DNA DNA DNA DNA 5 DNA 3 dNTP DNA IafRa PCR EL LLL 2 PCR 7 CY 1 DNA DNA DNA 94C 98 DNA
43. 8 lt 3 2 1 160 ml G 1XTAE 160mL 1 6 g 6 20 60 1 3
44. PCR 3 5 exonuclease x 5 3 exonuclease x 3 dA Taq PrimeSTAR 2 ov Tks Gflex M DNA 5 3 DNA 3 5 exonuclease co e DNA 3 5 exonuclease Pol 1 2 PCR e DNA DNA 5 3 5 3 3 5 exonuclease q
45. Iaf4H4 1 HHL Takaha maeofe PCR PCR Polymerase Chain Reaction DNA DNA 100 PCR Polymerase Chain Reaction DNA 100 PCR PCR DNA PCR
46. bromide EtBr 10mg mL DNA DNA SYBR DNA MVW Takaha Pramome Lo F a A gt
47. 0 w v 10 100 6 0 w v 300 7 000 1 2 w v zo0 4000 15 WA 100 3 000 2 0 w v 150 800 2 0 w v 100 600 3 0 w v Size Range bp SeaPlaque 50 250 4 0 w v 500 25 000 0 75 w v 300 20 000 1 0 w v 200 12 000 1 25 w v WN 1 596 0 1xTAE buffer 50 2 000 2 0 w v 1T lahaha PCR 5 PCR 1 Pol I gw PCR DNA 2 PolI 5 3 exonuclease PCR 3 dA Taq EmeraldAmp e 3 5 _exonuclease PCR PrimeSTAR Tks Gflex 3 5 exonuclease 5 3 5 3 exonuclease 5 3
48. 5 3 3 CAGT 37 3 mC AG T emm 5 eg cs CA 3 Ve 57 CAGT 5 Gm 3 5 T 3 T ee 5 a G CA er TGAC a D GC GC 40 60 G AT C Tm 5 em G TCA CAGT 3 LL GCC AT Tm 55 60 Tm Tm Tm melting temperature DNA DNA DNA
49. IdfaR4 PCR DNA DNA PCR PCR PCR 3 gt 1 DNA 0 4 2 PCR P6 0 66 6 3 PCR 0 7 4 9 5 PCR 12 gt 1 PCR 1 4 2 POR 1 7 3 2 4 PCR 2 6 5 2 7 wm 3 0
50. PCR Et N NN 50 PCR DNA DNA 20mer mer dNTP 20 PCR e 3 3
51. revention Kit 6088 200 42 000 30 L18 Gradient Thermocycler This product Model TP600 is licensed under U S Patent Nos 5 525 300 5 779 981 6 054 263 6 962 821 and corresponding foreign counterparts thereto L 45 HeatedCover This product is covered by the claims of U S Patent 5 552 580 and their corresponding foreign counterpart patent claims
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