临床 PCR 检验标本的采集、处理、保存及核酸提取方法

PCR

199768.2%13.3%18.5%200761.9%

PCR

PCR

PCR

,,

,RNAGITC()DNAEDTA,

DNARNAEDTA(PCR)6(2)12-20-70

,,EDTA-Na2,DNA,4,RNARNA

,,,,-70.

,DNA,,1 mol/L NaOHPCR ,-70

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)

1:0.1 mol/L NaOH50gK0.05% Triton X-100

GuSCNDNAPCR-alpha-casein

PCR,,,,510,,,DNA,-70.

,(),DNA,23,DNA,,,,,,DNA;,-70

,,,,,

HBV DNAHCV RNAPCR

DNA 50 mmol/L Tris-HCl10 mmol/L EDTA, 2% Triton X-100, 4 mol/L GITC, 0.3 mol/L 425-600um

,,K50%,,1cm,,,50%.,46,,KDNA

DNARNA PCR

PCR : :UV

MLVTAQ DNADNARNA

g0.1U,100%

I 1~3 U/g(5 mM Tris pH7.5, 1 mM CaCl2, 40 U RNase 25 2)

PCR,1PCRDNAHeminPCR21%(V/V)Taq

IgG IgGDNADNADNA

PCR,IgGPCR

NaOHTaq DNADNANaOHDNA

0.6% (wt/vol)BSATaq DNA2%vol/vol0.2%BSATaq DNA1020DNAT432gp32BSA

,, (Triton-100)(K)

DNA-

RNA,RNARNaseRNase,

RNaseRNaseRNA,RNA

RNA,RNase,RNA. RNase,1808,0.1%(DEPC)RNA,DEPCRNaseDEPC3721001515DEPC,DEPCRNA

RNA RNA,RNA,,RNase,0.1%DEPC3712,1001515

,DEPC,Tris,,TrisRNase

RNARNase RNase RNA,RNA, RNA

RNA

K-

-

-

SPIN COLUMN Se NaI Immobilon-P

SPIN COLUMN123

Se

NaI

m2000 DNA/RNAPCR 96

SPRI-TE1-10 Vidiera NsP:96

QIAGEN BioRobot MDx Workstation96

MagNA Pure LC 2.0 System

A260A260/A280/

1

1 2 PCR

PCR2-3cm2cm

34 RNAase

5 EDTAPCR

2 3 RNAchao tropic4mol/L(Guanidinium isothiocyanae,GITC),

5mol/LGITCRNase4mol/LRNaseRNAGITC7RNARNAEDTA70

DNA10mmol/LTris,lmmol/LEDTA(pH7.5~8.0)4RNA8020