临床 pcr 检验标本的采集、处理、保存及核酸提取方法
Post on 19-Mar-2016
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PCR
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199768.2%13.3%18.5%200761.9%
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PCR
PCR
PCR
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,,
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,RNAGITC()DNAEDTA,
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DNARNAEDTA(PCR)6(2)12-20-70
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,,EDTA-Na2,DNA,4,RNARNA
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,,,,-70.
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,DNA,,1 mol/L NaOHPCR ,-70
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1Universal sample processing, USP 4-6 mol/LGuHCl 50 mmol/L Tris-HCl, pH7.5 25 mmol/L EDTA 0.5%Sarkosyl 0.1~0.2 mol/L -20.05%Tween 80310%Chelex-100( 0.03% Triton X-1000.3% Tween 20)
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1:0.1 mol/L NaOH50gK0.05% Triton X-100
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GuSCNDNAPCR-alpha-casein
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PCR,,,,510,,,DNA,-70.
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,(),DNA,23,DNA,,,,,,DNA;,-70
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,,,,,
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HBV DNAHCV RNAPCR
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DNA 50 mmol/L Tris-HCl10 mmol/L EDTA, 2% Triton X-100, 4 mol/L GITC, 0.3 mol/L 425-600um
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,,K50%,,1cm,,,50%.,46,,KDNA
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DNARNA PCR
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PCR : :UV
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MLVTAQ DNADNARNA
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g0.1U,100%
I 1~3 U/g(5 mM Tris pH7.5, 1 mM CaCl2, 40 U RNase 25 2)
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PCR,1PCRDNAHeminPCR21%(V/V)Taq
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IgG IgGDNADNADNA
PCR,IgGPCR
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NaOHTaq DNADNANaOHDNA
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0.6% (wt/vol)BSATaq DNA2%vol/vol0.2%BSATaq DNA1020DNAT432gp32BSA
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,, (Triton-100)(K)
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DNA-
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RNA,RNARNaseRNase,
RNaseRNaseRNA,RNA
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RNA,RNase,RNA. RNase,1808,0.1%(DEPC)RNA,DEPCRNaseDEPC3721001515DEPC,DEPCRNA
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RNA RNA,RNA,,RNase,0.1%DEPC3712,1001515
,DEPC,Tris,,TrisRNase
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RNARNase RNase RNA,RNA, RNA
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RNA
K-
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SPIN COLUMN Se NaI Immobilon-P
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SPIN COLUMN123
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Se
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NaI
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m2000 DNA/RNAPCR 96
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SPRI-TE1-10 Vidiera NsP:96
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QIAGEN BioRobot MDx Workstation96
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MagNA Pure LC 2.0 System
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A260A260/A280/
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1
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1 2 PCR
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PCR2-3cm2cm
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34 RNAase
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5 EDTAPCR
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2 3 RNAchao tropic4mol/L(Guanidinium isothiocyanae,GITC),
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5mol/LGITCRNase4mol/LRNaseRNAGITC7RNARNAEDTA70
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DNA10mmol/LTris,lmmol/LEDTA(pH7.5~8.0)4RNA8020
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