KS J 4207-2011 Spectrophotometric and fluorometric analysis methods for GMO quantification《为转基因变形生物体(GMO)定量分析所采用的分光分析法及荧光分析法》.pdf

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3、J 0002 . 4 4.1 DNA 4.2 DNA 4.3 GMO( ) 5 5.1 DNA 5.1.1 5.1.1.1 ( ) , ( ) . KS J 4207:2011 2 5.1.1.2 . 5.1.1.3 3 % . 3 (m) (s) . 5.1.2 5.1.2.1 . 5.1.2.1.1 . 5.1.2.1.2 , . 5.1.2.2 (baseline) . 5.1.2.2.1 DNA . , DNA Tris DNA . 5.1.2.2.2 DNA 260 nm , . , DNA . 5.1.2.2.3 2 (drift) . 5.1.2.3 DNA 220 330 nm

4、 . DNA 230 nm, 260 nm, 280 nm 320 nm DNA . 5.1.2.3.1 DNA , DNA . . 5.1.2.3.2 / 3 . DNA . 5.1.2.3.3 KS J 4207:2011 3 (： ) . 5.1.2.3.4 DNA . , 400 g/mL DNA DNA (： ) . 5.1.2.4 DNA 150 200 g/mL DNA (serial dilution) . 3 . 5.1.2.4.1 DNA 260 nm , (least square method) (linear regression curve) . 5.1.2.4.2

5、 , . 5.1.2.5 DNA DNA DNA . 5.1.2.5.1 220 330 nm 260 nm 320 nm DNA 3 . 5.1.2.5.2 . 5.1.2.5.3 260 nm . 260 nm DNA , DNA DNA DNA . 5.1.2.5.4 5.1.2.5.2 5.1.2.5.3 , 260 nm (A260) 320 nm (A320) DNA (A260,cal) 1. , DNA A260,cal (A260 A320) DNA . 5.1.2.5.4.1 (double stranded) DNA A260,cal A260,cal 50 g/(mLO

6、D) , 50 g/(mLOD). 5.1.2.5.4.2 (single stranded) DNA A260,cal A260,cal 33 g/(mLOD) , 33 g/(mLOD). KS J 4207:2011 4 5.1.2.5.4.3 RNA A260,cal A260,cal 40 g/(mLOD) , 40 g/(mLOD). 5.1.2.5.4.4 DNA A260,cal (： 1/10 DNA 10 ). 5.1.2.6 DNA DNA (： , , , ) 1. , DNA . 5.1.2.6.1 230 nm, 260 nm 280 nm DNA 3 . 5.1.

7、2.6.2 . 5.1.2.6.3 . DNA , DNA DNA DNA . 5.1.2.6.4 5.1.2.6.2 5.1.2.6.3 , 260 nm (A260) 230 nm (A230) A260 280 nm (A280) DNA . 5.1.2.6.4.1 A260/A230 0.8 DNA . (： ) . 5.1.2.6.4.2 A260/A280 1.7 2.0 DNA . ( ： ) . 5.1.2.7 DNA . 5.1.2.7.1 (spin column silica membrane) DNA DNA , 260 nm . 5.1.2.7.2 DNA (CTAB

8、) 260 nm CTAB DNA DNA 260 nm DNA . 5.2 DNA 5.2.1 KS J 4207:2011 5 5.2.1.1 Quant-iT PicogreenTM DNA (Invitrogen ) DNA . 5.2.1.2 , . 5.2.1.3 DNA . 5.2.1.4 5 % . 3 (m) (s) . 5.2.2 5.2.2.1 . 5.2.2.1.1 . 5.2.2.1.2 , . 5.2.2.2 DNA , . DNA . 5.2.2.2.1 DNA 3 , 5 1 . 5.2.2.2.2 / 3 . DNA . 5.2.2.2.3 ( : ) . 5

9、.2.2.2.4 DNA . , 400 g/mL DNA / DNA/ ( : ) . 5.2.2.2.5 DNA (incubation) , KS J 4207:2011 6 . 5.2.2.3 DNA DNA . 5.2.2.3.1 DNA DNA DNA (calibration curve) . 5.2.2.3.2 DNA . DNA DNA . 3 . 5.2.2.3.3 DNA . DNA , DNA . 5.2.2.3.4 DNA DNA . 5.3 GMO( ) 5.3.1 5.3.1.1 GMO( ) DNA (dilution series) , DNA . 5.3.1

10、.2 DNA (dilution series) DNA . 5.3.2 5.3.2.1 DNA DNA (“ GM 1” ). 5.3.2.2 “ GM 1” 1/2 (“ GM 2” ). 5.3.2.3 “ GM 2” 1/4 (“ GM 3” ). 5.3.2.4 DNA 3 . KS J 4207:2011 7 5.3.2.5 DNA 3 (cycle number) DNA DNA DNA . 5.3.2.6 GM 1, 2, 3 DNA . 5.3.2.7 1 DNA . 5.3.2.7.1 GM 3： GM 2： GM 1 1:4:8 GM 1 DNA . 5.3.2.7.2

11、GM 1/ GM 2 2 (“2” ) “ GM 1” “ GM 2” GMO( ) . 5.3.2.7.3 GM 2/ GM 3 4 (“4” ) “ GM 2” GM 3 DNA . 5.3.2.7.4 , GM 2/ GM 3 4 (“4” ) “ GM 3” GMO( ) DNA GM 2 DNA . 5.3.2.8 DNA DNA DNA DNA(calibrator DNA) DNA . KS J 4207:2011 8 1 https:/www.matcmadison.edu/biotech/resources/methods/spectro/spectro.htm, The M

12、ATC Biotechnology Project 2 KS J ISO 21569, 3 KS J ISO 21570, 4 KS J ISO 21571, 5 ISO/TS 21098： 2005, Foodstuffs Nucleic acid based methods of analysis of genetically modified organisms and derived products Information to be supplied and procedure for the addition of methods to ISO 21569, ISO 21570 or ISO 21571 (GMO) 153787 1 92 3(13) (02)26240114 (02)262401489 http:/ Korean Agency for Technology and Standards http:/www.kats.go.kr KS J 4207:2011 KSKSKS SKSKS KSKS SKS KS SKS KSKS SKSKS KSKSKS Spectrophotometric and fluorometric analysis methods for GMO quantification ICS 67.050