CN103667254B - 目标基因片段的富集和检测方法 - Google Patents
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Abstract
Description
反应材料 | 体积 |
纯化后cDNA/DNA样本库 | 50微升 |
磷酸化反应缓冲液 | 10微升 |
脱氧碱基混合物dNTP(每种10mM) | 4微升 |
T4DNA聚合酶 | 5微升 |
Klenow大肠杆菌聚合酶片段 | 1微升 |
T4多聚核苷酸激酶 | 5微升 |
无核酸酶水 | 总体积补至100微升 |
反应材料 | 体积 |
cDNA/DNA样本库 | 约30微升 |
10X Klenow大肠杆菌聚合酶缓冲液 | 5微升 |
脱氧碱基dATP(1mM) | 10微升 |
Klenow大肠杆菌聚合酶片段 | 3微升 |
无核酸酶水 | 总体积补至50微升 |
反应材料 | 体积 |
cDNA/DNA样本库 | 约15微升 |
2XT4DNA连接酶缓冲液 | 5微升 |
DNA两端接头 | 6微升 |
T4DNA连接酶 | 3微升 |
无核酸酶水 | 总体积补至50微升 |
ABL1 | LCX | ELK4 | RAD51L1 | CBLB | NFE2L2 | GNA11 | TCL6 |
ABL2 | LHFP | ELKS | RAF1 | CBLC | NFIB | GNAQ | TET2 |
ACSL3 | LIFR | ELL | RALGDS | CCDC6 | NFKB2 | GNAS | TFE3 |
AF15Q14 | LMO1 | ELN | RANBP17 | CCNB1IP1 | NIN | GOLGA5 | TFEB |
AF1Q | LMO2 | EML4 | RAP1GDS1 | CCND1 | NKX2-1 | GOPC | TFG |
AF3p21 | LPP | EP300 | RARA | CCND2 | NONO | GPC3 | TFPT |
AF5q31 | LRIG3 | EPS15 | RB1 | CCND3 | NOTCH1 | GPHN | TFRC |
AKAP9 | IYL1 | ERBB2 | RBM15 | CCNE1 | NOTCH2 | GRAF | THRAP3 |
AKT1 | MADH4 | ERCC1 | RECQL4 | CD273 | NPM1 | H3F3A | TIF1 |
AKT2 | MAF | ERCC2 | REL | CD274 | NR4A3 | HCMOGT-1 | TLX1 |
ALDH2 | MAFB | ERCC3 | RET | CD74 | NRAS | HEAB | TLX3 |
ALK | MALT1 | ERCC4 | ROS1 | CD79A | NSD1 | HERPUD1 | TMPRSS2 |
’ALOI7 | MAML2 | ERCC5 | RPL22 | CD79B | NTRK1 | HEY1 | TNFAIP3 |
APC | MAP2K4 | ERG | RPN1 | CDH1 | NTRK3 | HIP1 | TNFRSF14 |
ARHGEF12 | MDM2 | ETV1 | RUNDC2A | CDH11 | NUMA1 | HIST1H4I | TNFRSF17 |
ARHH | MDM4 | ETV4 | RUNX1 | CDK12 | NUP214 | HLF | TNFRSF6 |
ARID1A | MDSI | ETV5 | RUNX2 | CDK4 | NUP98 | HLXB9 | top1 |
ARID2 | MDS2 | ETV6 | RUNXBP2 | CDK6 | OLIG2 | HMGA1 | TP53 |
ARNT | MECT1 | EVI1 | SBDS | CDKN2A | OMD | HMGA2 | TPM3 |
ASPSCR1 | MED12 | EWSR1 | SDC4 | CDKN2a(p14) | P2RY8 | HNRNPA2B1 | TPM4 |
ASXL1 | MEN1 | EXT1 | SDH5 | CDKN2B | PAFAH1B2 | HOOK3 | TPR |
ATF1 | MET | EXT2 | SDHB | CDKN2C | PALB2 | HOXA11 | TRIM27 |
ATIC | MITF | EZH2 | SDHC | CDX2 | PAX3 | HOXA13 | TRIM33 |
ATM | MKL1 | EZR | SDHD | CEBPA | PAX5 | HOXA9 | TRIP11 |
ATRX | MLF1 | FACL6 | SMAD4 | CEP1 | PAX7 | HOXC11 | TSC1 |
BAP1 | MLH1 | FAM22A | SEP6 | CHCHD7 | PAX8 | HOXC13 | TSC2 |
BCL10 | MLL | FAM22B | SET | CHEK2 | PBRM1 | HOXD11 | TSHR |
BCL11A | MLL2 | FAM46C | SETD2 | CHIC2 | PBX1 | HOXD13 | TTL |
BCL11B | MLL3 | FANCA | SF3B1 | CHN1 | PCM1 | HRAS | U2AF1 |
BCL2 | MLLT1 | FANCC | SFPQ | CIC | PCSK7 | HRPT2 | USP6 |
BCL3 | MLLT10 | FANCD2 | SFRS3 | CIITA | PDE4DIP | HSPCA | VHL |
BCL5 | MLLT2 | FANCE | SH3GL1 | CLTC | PDGFB | HSPCB | VTI1A |
BCL6 | MLLT3 | FANCF | SIL | CLTCL1 | PDGFRA | IDH1 | WAS |
BCL7A | MLLT4 | FANCG | SLC34A2 | CMKOR1 | PDGFRB | IDH2 | WHSC1 |
BCL9 | MLLT6 | FBXO11 | SLC45A3 | COL1A1 | PER1 | IKZF1 | WHSC1L1 |
BCOR | MLLT7 | FBXW7 | SMARCA4 | COPEB | PHF6 | IL2 | WIF1 |
BCR | MN1 | FCGR2B | SMARCB1 | COX6C | PHOX2B | IL21R | WNT1 |
BHD | MPL | FEV | SMO | CREB1 | PICALM | IL6ST | WRN |
BIRC3 | MSF | FGFR1 | SOCS1 | CREB3L1 | PIK3CA | IL7R | WT1 |
BLM | MSH2 | FGFR1OP | SOX2 | CREB3L2 | PIK3R1 | IRF4 | WTX |
BMPR1A | MSH6 | FGFR2 | SRGAP3 | CREBBP | PIM1 | IRTA1 | WWTR1 |
BRAF | MSI2 | FGFR3 | SRSF2 | CRLF2 | PLAG1 | ITK | XPA |
BRCA1 | MSN | FH | SS18 | CRTC3 | PML | JAK1 | XPC |
BRCA2 | MTCP1 | FHIT | SS18L1 | CTNNB1 | PMS1 | JAK2 | XPO1 |
BRD3 | MUC1 | FIP1L1 | SSH3BP1 | CYLD | PMS2 | JAK3 | XRCC1 |
BRD4 | MUTYH | FLI1 | SSX1 | D10S170 | PMX1 | JAZF1 | YWHAE |
BRIP1 | MYB | FLJ27352 | SSX2 | DAXX | PNUTL1 | JUN | ZNF145 |
BTG1 | MYC | FLT3 | SSX4 | DDB2 | POU2AF1 | KDM5A | ZNF198 |
BUB1B | MYCL1 | FNBP1 | STK11 | DDIT3 | POU5F1 | KDM5C | ZNF278 |
C12orf9 | MYCN | FOXL2 | STL | DDX10 | PPARG | KDM6A | ZNF331 |
C15orf21 | MYD88 | FOXO1A | SUFU | DDX5 | PPP2R1A | KDR | ZNF384 |
C15orf55 | MYH11 | FOXO3A | SUZ12 | DDX6 | PRCC | KIAA1549 | ZNF521 |
C16orf75 | MYH9 | FOXP1 | SYK | DEK | PRDM1 | KIF5B | ZNF9 |
C2orf44 | MYST4 | FSTL3 | TAF15 | DICER1 | PRDM16 | KIT | ZRSR2 |
CAMTA1 | NACA | FUBP1 | TAL1 | DNM2 | PRF1 | KLK2 | ATP6V1F |
CANT1 | NBS1 | FUS | TAL2 | DNMT3A | PRKAR1A | KRAS | BCAT2 |
CARD11 | NCOA1 | FVT1 | TCEA1 | DUX4 | PRO1073 | KTN1 | LAMP1 |
CARS | NCOA2 | GAS7 | TCF1 | EBF1 | PSIP2 | LAF4 | PDLA6 |
CBFA2T1 | NCOA4 | GATA1 | TCF12 | ECT2L | PTCH | LASP1 | PSMC1 |
CBFA2T3 | NDRG1 | GATA2 | TCF3 | EGFR | PTEN | LCK | VDAC2 |
CBFB | NF1 | GATA3 | TCF7L2 | EIF4A2 | PTPN11 | LCP1 | WARS |
CBL | NF2 | GMPS | TCL1A | ELF4 | RAB5EP |
反应材料 | 体积 |
富集cDNA/DNA样本库 | 约30微升 |
10X高准确率超保真DNA聚合酶缓冲液 | 5微升 |
高准确率超保真DNA聚合酶 | 1微升 |
正引物 | 1微升 |
反引物 | 1微升 |
无核酸酶水 | 总体积补至50微升 |
反应材料 | 体积 |
纯化后cDNA/DNA样本库 | 50微升 |
磷酸化反应缓冲液 | 10微升 |
脱氧碱基混合物dNTP(每种10mM) | 4微升 |
T4DNA聚合酶 | 5微升 |
Klenow大肠杆菌聚合酶片段 | 1微升 |
T4多聚核苷酸激酶 | 5微升 |
无核酸酶水 | 总体积补至100微升 |
反应材料 | 体积 |
cDNA/DNA样本库 | 约30微升 |
10X Klenow大肠杆菌聚合酶缓冲液 | 5微升 |
脱氧碱基dATP(1mM) | 10微升 |
Klenow大肠杆菌聚合酶片段 | 3微升 |
无核酸酶水 | 总体积补至50微升 |
反应材料 | 体积 |
cDNA/DNA样本库 | 约15微升 |
2XT4DNA连接酶缓冲液 | 5微升 |
DNA两端接头 | 6微升 |
T4DNA连接酶 | 3微升 |
无核酸酶水 | 总体积补至50微升 |
Ct值第一次重复 | Ct值第二次重复 | Ct值第三次重复 | Ct均值 | Ct差 | 富集倍数 | |
未富集样本 | 36.97 | 37.05 | 37.02 | 37.01 | 0 | 1 |
已优化探针 | 23.79 | 23.84 | 23.75 | 23.79 | 13.22 | 9546.50 |
反应材料 | 体积 |
富集cDNA/DNA样本库 | 约30微升 |
10X高准确率超保真DNA聚合酶缓冲液 | 5微升 |
高准确率超保真DNA聚合酶 | 1微升 |
正引物 | 1微升 |
反引物 | 1微升 |
无核酸酶水 | 总体积补至50微升 |
Ct值第一次重复 | Ct值第二次重复 | Ct值第三次重复 | Ct均值 | Ct差 | 富集倍数 | |
未富集样本 | 36.56 | 36.65 | 36.94 | 36.72 | 0 | 1.0 |
已优化探针 | 23.36 | 23.39 | 23.34 | 23.36 | 13.35 | 10430.4 |
Ct值第一次重复 | Ct值第二次重复 | Ct值第三次重复 | Ct均值 | Ct差 | 富集倍数 | |
未富集样本 | 34.96 | 34.98 | 35.15 | 35.03 | 0 | 1 |
已优化探针 | 22.01 | 22.04 | 21.97 | 22.01 | 13.02 | 8324.2 |
Claims (10)
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