DB21T 3084-2018 羽絨制品中鵝絨的鑒別PCR法和實(shí)時(shí)熒光PCR法　

DB21IdentificationofGooseDownFiberinFeatherandDownProducts-PCRmethodandreal-timefluorescenc遼寧省市場(chǎng)監(jiān)督管理局發(fā)布I請(qǐng)注意本文件的某些內(nèi)容可能涉及專利，本文件的發(fā)布機(jī)構(gòu)不承擔(dān)識(shí)別這些專利的責(zé)任。1羽絨制品中鵝絨的鑒別PCR法和實(shí)時(shí)熒光PCR法GB/T6682分析實(shí)驗(yàn)室用水規(guī)格和3.1聚合酶鏈?zhǔn)椒磻?yīng)polymerasechainrea3.2實(shí)時(shí)熒光PCRreal-timefluorescencepolymerasechainr4縮略語bp：堿基對(duì)（basepair）DNA：脫氧核糖核酸（deoxyribonuleicacid）dNTP：脫氧核苷酸三磷酸（deoxyribonucleosidetriphospha26.2組織和毛發(fā)提取試劑盒TaKaRaMiniBESTUniversalGenomicDNAExtractionKitVer.5.0(T3），10.8將核酸純化柱安置于收集管上），進(jìn)行PCR、實(shí)時(shí)熒光PCR鑒別時(shí)設(shè)置陽性對(duì)照、陰性對(duì)照和空白對(duì)照。陽性對(duì)照：擴(kuò)增鵝5’-GCTACTAGCCATACACTAC-3’5’-GTTGGATTGTCTACTGAGA-3’4并加DNAmarker點(diǎn)樣以判斷PCR產(chǎn)物的片段大小。電壓大小根據(jù)電泳鵝5’-GCTACTAGCCATACACTAC-3’5’-GTTGGATTGTCTACTGAGA-3’5’-（FAM）CGCAGACACTTCACTCGCCT（Eclipse）-3’反應(yīng)結(jié)束后，應(yīng)設(shè)置無效基線范圍，基線范圍選擇在3~15個(gè)循環(huán)5對(duì)照和空白對(duì)照結(jié)果正常者，則可判定該樣品檢出鵝源性6PremixExTaq（ProbeqPCR2×C7擴(kuò)增產(chǎn)物序列Attcctagccatgcactactcaccagacgcctcaaccgccttttcatcaatcgcccacatcactcgagacgtaaattatggctgaatcatccgctaccttcacgccaatggcgcctcaatattctttatctgcctcttcctacacatcgggcgaggcctatattactctctactcagaaacctgaaacatcggcattatcctcctgcttgcaactatagcaacagccttcataggctatgtccaaatatcattctgaggggccacagtaattacaaacttactatccgccatcc