15-07 - PCR- (Code No. FIK-101) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A4849K
[1] 1 [2] 2 [3] 2 [4] 2 [5] 2 [6] 4 [7] 6 [8] 7 [9] 8 0
[1] 50 50 1
[2] 2x PCR Master Mix 1ml 5-20 10x Primer Mix 1ml 1-20 UNG(Uracil-DNA Glycosylase 200 l 1-20 [3] 100bp 1.5ml 0.2mlPCR 20ml 1.5ml 95 12,000rpm MultiNA MultiNA DNA-1000 SYBR Gold Code:DNA-035 3 SYBR Molecular Probes Inc. [4] inva VT1, VT2, VT2vha, VT2vhb, VT2vp1 ipah ipah [5] PCR PCR 2
PCR PCR dttp dutp DNA UNG UNG dutp PCR dttp DNA UNG PCR UNG UNG UNG 10 4 10 6 / ( /) ipah 10 4 inva VT 10 6 InvA VT ipah 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 0 2 10 2 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 3 10 4 4 10 6 5 10 8 InvA VT ipah 3
[6] A, B A 1) 1.5ml 50μl 2) 1-2 3) 10μl 50 B 1) 20ml 2.5ml 2) 2-3 3) 2-3 4) 50 5) 2-3 6) 0.5ml 1.5ml 1) 1.5ml 95 5min 2) 12,000rpm X 3 3) 2μl PCR 4 PCR PCR 1) 2) 3) 4) 18μl 5.6 μl 2x PCR Master Mix 10.0 μl 10x Primer Mix 2.0 μl UNG 0.4 μl 2.0 μl 20.0 μl 4
PCR UNG : 20, 10 95, 2 : 94, 10 : 57, 30 10 : 68, 30 : 94, 10 : 55, 30 15 : 68, 30 : 94, 10 : 53, 30 20 : 68, 30 4120 MultiNA PCR DNA 100bp DNA Ladder DNA-035 3 MultiNA 685 735bp 5 mv 610 655bp 7 mv 430 480bp 2mV 260 290bp 2 mv 5
(1) 2 (2) [7] 6
[8] O157 100 1000 O157 cfu/g) 0 7x10 2 7x10 3 7x10 4 7x10 5 7x10 6 PCR cfu/g) 0 7 x10 2 7 x10 3 7 x10 4 7 x10 5 7 x10 6 PCR cfu/g) 0 4 x10 2 4 x10 3 4 x10 4 4 x10 5 4 x10 6 PCR 50 O157 cfu/g) 1 10 20 40 50 60 70 80 90 100 7x10 2 7x10 3 7x10 4 7x10 2 7x10 3 7x10 4 4 x10 2 4 x10 3 4 x10 4 7
[9] 10 UV inva, VT, ipah 10 UV 8
9
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