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マアジ及びニシマアジの 魚種判別マニュアル

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Contents

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5. DNA Proteinase K RNase A DNA DNA DNA NADH dehydrogenase subunits 5 ND5 PCR PCR DNA bO ND5 D 128 R
6. EcoR I TOYOBO H Buffer PCR DNeasy Blood amp Tissue kit QIAGEN DNA 1 5 mL 25 mg 100 1 000 H L 180 H L Buffer ATL 10 100 u L 20 p L Proteinase K Kit 15 55 C 2 0 5 10 H L 100 mg mL RNase A 4 0 H L 15 2 15 100 1 000 u L 200 u Buffer AL 15 70 90110
7. PCR PCR 02 mL PCR DNA PCR PCR 0 2 mL 45 L PCR PCR DNA 5uL DNA 5uL DNA 5 uL DNA 5H L DNA PCR PCR PCR PCR PCR PCR JAS PCR 3 360bp DNA PCR DNA PCR PCR PCR Hinf 1 EcoR I 02 mL PCR 0 2 mL lOu L PCR PCR 10H L DNA PCR lOu L Hinf I EcoR I 37 C 1 JAS PCR 3 96 wlv Hinf I Ec
8. 0 5 mL 37 C PCR DNA DNeasy Blood amp Tissue Kit DNeasy 96 Blood amp Tissue Kit QIAGEN RNase A 100 mg mL PCR DNA AmpliTaq Gold Applied Biosystems dNTP 2 mmol L each Gold MgCl 25 mmol L AmpliTaq Gold PCR 100 PCR buffer II AmpliTaq Gold DNA DNeasy Blood amp Tissue kit QIAGEN DNA DNA Lt1 ND5 Htl ND5 Ltl NDS 0 Htl ND5 JAS PCR
9. Proteinase 180 u L K Buffer ATL working solution Buffer ATL 20 u L 00 1 000 H L 200 H L 00 1 000 u L DNeasy mini column 6 000 x g 1 2 mL 100 1 000 u Buffer AW1 500 u L 6 000 x g 1 2 mL 100 1 000 H L Buffer AW2 500 u L 12 000 x g 3 2 mL 100 1 000 u L 200 H L 1 6 000 x g 1 DNA 1 DNA PCR DNA 20 C DNeasy 96 Tissue Kit QIAGEN DNA Collection micro tube 25 mg proteinase K Kit
10. Collection microtube 100 1 000 H L 8 12 Buffer AL E 410 U L Collection microtube Collection microtube 15 3 000 X g DNeasy 96 Plate S Block Collection microtube 100 1 000 H L 8 12 500 H L DNeasy 96 Plate AirPore Tape Sheet DNeasy 96 Plate DNeasy 96 Plate 5 635 x g 10 AirPore Tape Sheet 100 1 000 u L 8 12 Buffer AW1 500 H L AirPore Tape Sheet DNeasy 96 Plate DNeasy 96 Plate 5 635 x gl 5 AirPore Tape Sheet 100 1 000 L 8 12 Buffer AW2 500 U L AirPore Tape Sheet DNeasy 96 Plate DNeasy 96 Plate 5 635 x gl 5 AirPore Tape Sheet DNeasy 96 Plate Elution Microtube RS 70 15 100 1 000 H L 8 12 70 C 100 u L AirPore Tape Sheet DNeasy 96 Plate 1 5 635 X gl 1 DNA AirPore Tape Sheet 1 DNA PCR DNA DNA PCR 121 CH 15 PCR DNA PCR PCR DNA PCR DNA DNA PCR
11. DNeasy 96 Plate membrane PCR JAS PCR PCR JAS PCR PCR JAS PCR
12. RFLP RFLP RFLP RFLP Lt1 ND5 Ht1 ND5 DNA ND5 Lt1 ND5 Ht1 ND5 DNA 37 15 35 14 2 1 RFLP Lt1 ND5 Ht1 ND5 DNA DNA RFLP PCR DNA PCR Yasuharu T Takami M Michiaki Y Complete mitochondrial DNA sequence of Atlantic horse mackerel Trachurus trachurus and molecular identification of two commercilly important species trachurus and T japonicus using PCR RFLP Fisheries Science2006 72 1054 1065 Lt1 ND5
13. 514 12 31 12 16 31 516 12 50 31 Trachurus japonicus trachurus T DNA DNA
14. Proteinase K Buffer ATL working solution Collection microtube Collection microtube 100 1 000 H L 8 12 Collection microtube cap 200 u L Kit Collection microtube 3 000 x g 55 C 2 3 000 x g Collection microtube 100 mg mL Collection microtube 15 3 000 x g
15. 12 DOO0d 000000 000000000 000000000000 0000000000000 175 pog00000000 14 1680000 10000000 000000000 0000000000 0 000000000000000000000000000000000000000000 0 000000000000000000000000000000000000000000 0 0000000 DNase PCR 15 121 C DNA 0 2 mL
16. Hinf I 12 U u L 10x H buffe r PCR 20 2 5 U tube 1x ML 2 EcoRI EcoR I 8 L 10x H buffer PCR 2 5 U tube 1x Trachurus trachurus Trachurus japonicus 152 Ltl NDS 10 20 30 40 P gt C A C T A A G T TP P G ACT G T CCOA ACATCEAGECLTAAAATICGACEA 50 60 70 80 l l l l C T A G P C T A T A T T T CA C C GC CS A p CG GO COCO T C TP A COS TC A A T 90 EcoR I 100 110 120 TGRTCCATTCTCICAATTCIGCATCATGCRTACATGCACGCAG ma Oe ccc 130 140 150 160
17. 0000 uu 5 30 LU Lt NO DO OO DNA Lt ND5 360 bp CACTAACCTTTGATGTCAACAT NADH dehydrogenase subunits 5 NDS Ht1 ND5 TTGCTATTCATGCTATGGCGA ND 0000 PCR PCR DNA DNA X DNA DNA DNA 4 PCR tube 30 75 L Gold 5 U u L 0 25 u L 1 25 U tube 10x PCR buffer II 5 ML 1x dNTP 2 mmol L each 5 ML 200 u mol L each MgCl 25 mmol L 3 ML 1 5 mmol L 5 50 mol L 05 HL 0 5 u mol L 3 50 H mol L 05 HL 0 5 H mol L DNA 5 uL 50 UL PCR 4 C 10 PCR Hinf I EcoR I DNA X X DNA DNA DNA X X 1 HinfI
18. Go 4 G 170 180 190 200 l l l CCTCATTGCUCATOGATTATTUCTAGTCACTGCAAACAACATOG Eos eR wa cw CES Ne e eodem 210 220 230 Hinf 1 240 Q e au 250 260 270 280 l l l l CTTTCCTYCTAATTGGUTGATGATACGGGCUGAACAGATGU 290 300 310 BET 320 C A ACACUGUCCGUCCTAUCAAGCAGTTUCTCTACAACCGGOGITILU 330 340 350 Ht1 ND5 360 GGEOTGACATCGGACTAATTTTCGCGCCCATAGCCATGAATAGCAA T Ltl ND5 Ht1 NDS5 40 40 80 80 120 120 160 160 200 200 240 240 280 280 320 320 360 360 R A G Y C T 13 joononoononnononnonnononnonnnnennoneon joononoononnononnonnononnonnnnennoneon 2I000000000000000000000000000000000000000000000000000 00000000000000000 0 s ooononoononnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn 00000000000000000000000000000000000000000000000000 00000 00000000
19. 1 5 mL JAS DNA ii PCR DNA 0 0056 mS m 25 C TE
20. RNase A Collection microtube cap 40H L 0 5 10 u L 12 Collection microtube 15 2 3 000 x g
21. 190 10 191 uje 000000 25 000000 0000000000 ii iii PCR PCR PCR Lt1 ND5 Ht1 NDS5 514 25
22. Ht1 ND5 PCR DNA
23. AA ese 9jnnnunnunnnnnunnunnunnnnnnnnunnnnnne 1410000000000000000000000000000000000000000000000000000 0000000000000 000000000000 1610000000000000000000000000000000000000000000000000000 000000000000 Ll 2 0 14 2003 30 7 18 6 18 3 1 7 0 8 0 2 0 2 0 1 70 6 9 6 7 7 7 296 3 0 1 6 0 1 0 1 29 4 100 0 E um c d t A 2 soe mom A ws gt ws x RAKTITTI T a 000000000 3040 __ aml uw BEE m gt mua mun ao gt A mes Ts iss 0000000 0000000 0000000 000000060 0 0000 U 0 000 0 00 0 000000 00000000000
24. DNase ili JAS iv 2 DNA
25. 00000000000000 00000 00000 0000 00 0 0000000000000 00000 000 0000 16 17
26. NA rRNA 16S rRNA DNA DNA D bl 12S rRNA 16S rRNA DNA DNA Ltl ND5 Ht1 ND5 PCR Hinf I EcoR I PCR DNA DNA QIAGEN DNeasy Blood amp Tissue kit DNeasy 96 Blood amp Tissue kit RFLP DNA 1 5mL 2 0mL 12 000 x g 050 1 000 L 0 5 10 H 10 100 H 100 1 000 L 55 C 70 C PCR PCR GeneAmp PCR System 9700 Applied biosystems 050 1 000 L 0 5 10 u 10 100 H 100 1 000 u L 0 2 mL 1 5mL JAS JAS PCR 050 1 000 u L 0 5 10 u LO 10 100 100 1 000 u L
27. oR I

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