サバ属魚類の魚種判別マニュアル - 農林水産消費安全技術センター
Contents
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4. 100 1 000 u L 8 12 200 p L Proteinase K Buffer ATL working solution Collection microtube cap Kit Collection microtube Collection microtube 3 0
5. Hae Hinf TOYOBO PCR DNeasy Blood amp Tissue 01 QIAGEN DNA 1 5 mL 25 mg 100 1 000 L 180 u L Buffer ATL 10 100 u L 20 p LI Proteinase K Kit 15 55 C 2 55 C 0 5 10 u L 100 mg mL RNase 401 L 15 2 15 100 1 000 u L 200 u L Buffer AL 15 70 90110 100 1 000 u L 200 H L 15 100 1 000 H L DNeasy mini column
6. DNeasy 96 Plate S Block Collection microtube 100 1 000 H L 8 12 50011 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 L AirPore Tape Sheet DNeasy 96 Plate DNeasy 96 Plate 5 635 X g 5 AirPore Tape Sheet 100 1 000 Hu L 8 12 Buffer AW2 500 L AirPore Tape Sheet DNeasy 96 Plate DNeasy 96 Plate 5 635 X g 5 AirPore Tape Sheet DNeasy 96 Plate Elution Microtube RS 70 15 100 1 000 u L 8 12 70 C 100 u L AirPore Tape
7. DNase ili JAS iv 2 DNA DNA Proteinase
8. 0 2 mL 45 L PCR PCR DNA Su L DNA 51117 DNA 5 uL DNA Su L DNA PCR PCR PCR 4 PCR PCR PCR JAS PCR 3 PCR 505 bp DNA PCR PCR PCR PCR Hae Hinf I 0 2 mL 6 PCR 0 2 mL 10111 PCR 10H L DNA PCR 10 u L Hae 111 Hinf 1 37 00 15
9. 100 1 000 u L Buffer AW1 500 u L 6 000 x g 1 2 mL 100 1 000 u L Buffer AW2 500 u L 12 000 x g 3 2 mL 100 1 000 u L 200 11 L 1 6 000 x g 1 DNA 1 DNA PCR DNA 20 DNeasy 96 Blood amp Tissue QIAGEN DNA Collection micro tube 25 mg Proteinase K Buffer ATL working solution 20 LI proteinase Kit 180 p L Buffer ATL
10. JAS PCR 3 w v Hinf I PCR PCR 7 Hae Hinf I RFLP RFLP RFLP 7 RFLP LSs1 Leu HSs1 ND5 DNA PCR LSs1 Leu HSs1 ND5 PCR DNA 15 15 15 RFLP PCR DNA PCR 5 1999 Yasuharu T Takami M Michiaki Y Complete mitochondrial DNA seguence of Atlantic horse mackerel Trachurus trachurus and molecular identification of two commercilly important species T trachurus and T japonicus using PCR RFLP Fisheries Science2006 72 1054 1065 LSs1 Leu HSs1 ND5 PCR DNA
11. PCR RFLP DNA 31 Scomber japonicus 12 PCR 175 0000000000D0000000000D0000000000000900000D00000D 0D0D000000000000140160 D D0M00NN00DD00000000000000 0000000000D0000000000D00000D0000000000000D00000D 0000000000D00000000000D000000000000D00000000000D
12. 514 12 31 16 12 50 31 516 australasicus 5 213 1 12
13. DNA X X Hae tube 7 875 L Hae 20 UN L 0 125 L 2 5 U tube 10x M Buffer 2 1 PCR 10 HL 20 UL Hinf I tube 7 875 L Hint I 20 L 0 125 L 2 5 U tube 10x H Buffer 2 ul PCR 10 HL 20 ML RFLP Scomber scombrus Scomber japonicus Hae III Scomber australasicus zl 11111 211 00 0 11111711111 00 0 0000000 00 0 00000000000 12 LSs1 Leu 10 20 30 40 SS ATCCGCTGGTCITAGGAACCAGAAACTCOTITGGIGCcAAA TC 50 60 70 80 CAAGTAGCAGCTAATGACTTCCACCTCCGTAACAATAACA 90 100 110 120 A GAA GC GC TA AT CAT ThA T CO T T C T C CC CC T AGcCOGCGcG M ET 130 140 150 160 TATTCACAACCCTCTCCCCCOCC
14. DNeasy 96 Plate membrane
15. 6 000 x 1 2 mL
16. 0 2 mL 05 104 L PCR GeneAmp PCR System 9700 Applied biosystems 0 50 1 000 u L 10 100 u 100 1 000 u L 1 5mL JAS JAS 050 1 000 u L 05 104 L 10 100 u 100 1 000 L PCR 0 5 mL DNA DNeasy Blood
17. DNA PCR 523 DNA LSs1 Leu 505bp ATCCGCTGGTCTTAGGAACC DNA HSs1 ND5 tR
18. 000 0000000000000 000000000000 0000000000000 0000000000000 0DD00000000 0000000000 0000000000 0000000000 14 00000000000000 000000000000000 000000000000000 0000000000 0000000000
19. 0D00000000000000D scombrus DNase PCR 15 121 C DNA 0 2 mL 1 5 mL DNA JAS ii PCR 0 0056 mS m 25 C
20. DNA QIAGEN DNA DNeasy Blood amp Tissue kit RFLP DNeasy 96 Blood amp Tissue kit LSs1 Leu DNA 1 5mL 2 0mL 12 000 x g 0 50 1 000 u L 05 101 10 100 u 100 1 000 u L 55 C 70 C
21. Sheet DNeasy 96 Plate 1 5 635 X gl 1 DNA AirPore Tape Sheet 1 DNA PCR DNA DNA PCR 121 15 DNA PCR PCR DNA PCR DNA PCR PCR PCR 0 2 mL 3 PCR DNA PCR PCR
22. amp Tissue Kit RNase A 100 mg mL PCR DNeasy 96 Blood amp Tissue Kit QIAGEN PCR DNA AmpliTag Gold Applied Biosystems dNTP 2 mmol L each AmpliTag Gold MgCl 25 mmol L AmpliTaq Gold PCR 100 PCR buffer II AmpliTaq Gold JAS PCR
23. 00 X g Collection microtube 55 C 2 3 000 55 C Collection microtube 15 3 000 X g Collection microtube 0 5 10 u L 8 12 100 mg mL RNase 40H L Collection microtube cap Collection microtube 15 2 3 000 X g Collection microtube 100 1 000 u L 8 12 Buffer AL E 410 H L Collection microtube cap Collection microtube 15 3 000 x g
24. 11121 11111 190 10 191 0000000000010 000000 0000000000 ii 514 5 25
25. AGCCCUCAAGCCCCTAACIG 170 180 190 200 4 AAA AGCAGTCGCGCAAGTCAAGACCGCAGTTAAACTGGCATTT oA E aD 210 220 230 240 TICGOTTACCTTACTCCOUCTTATITECTOTTIrATAAACGAAG 9 A amp A ie 250 260 270 280 san 290 300 310 320 330 340 350 360 TALTEGAITAITITTAECIETCCAATTECLACTETACETEACAT 370 380 390 400 GATCAATCCTTGAGTTTGCTTCATGATACATGCACGCTGA Jona cT ITI EM EM qu 410 420 430 440 Aint I CCCCTACATGAACCGCTTCTTTAAATACCTCCTGGTCTTT E EL I lS 450 260 470 480 ET Qiu ey ue i es T SE 500 HSs1 ND5 TTCAACTATTTATCGGCTGAGAAGG LSs1 Leu HSs1 ND5 13 0000 4 17
26. NA Leu CUN ND5 PCR PCR DNA DNA X DNA DNA DNA PCR tube 30 75 U AmpliTaqIM Gold 5 1 0 25 L 1 25 U tube 10x PCR buffer I 5 HL 1 dNTP 2 mmol L each 5 ul 200 mol L each MgCl 25 mmol L 3 pL 1 5 mmol L 5 5011 mol L 05 UL 0 5 u mol L 3 5011 mol L 0 5 UL 0 5 u mol L DNA 5 mL 50 HL PCR 10 Hinf I DNA X DNA
27. RNase A DNA DNA tRNA Leu CUN dehydrogenase subunit NADH 5 ND5 PCR PCR DNA D 128 rRNA 165 rRNA 01 125 rRNA LSs1 Leu HSs1 ND5 PCR
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