CN1164260A - 用新的碱基配对方案减少非特异性杂交 - Google Patents
用新的碱基配对方案减少非特异性杂交 Download PDFInfo
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- CN1164260A CN1164260A CN95195880A CN95195880A CN1164260A CN 1164260 A CN1164260 A CN 1164260A CN 95195880 A CN95195880 A CN 95195880A CN 95195880 A CN95195880 A CN 95195880A CN 1164260 A CN1164260 A CN 1164260A
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- C12N2310/33—Chemical structure of the base
- C12N2310/336—Modified G
Abstract
Description
SEQIDNO: | 表1捕获扩展物库A |
1 | ATTGCGAATAATAATTTTTTCACGTTGAAAATCTTCTCTTGGAAAGAAAGTGAT |
2 | GAATTTCTTAAACAGCTTGATACCGATAGTTGTTCTCTTGGAAAGAAAGTGAT |
3 | ATTGTATCGGTTTATCAGCTTGCTTTCGAGGTTTCTCTTGGAAAGAAAGTGAT |
4 | CCGCTTTTGCGGGATCGTCACCTTCTC TTGGAAAGAAAGTGAT |
5 | GCTGAGGCTTGCAGGGAGTTAAAGG TTCTCTTGGAAAGAAAGTGAT |
6 | ATGAGGAAGTTTCCATTAAACGGGT TTCTCTTGGAAAGAAAGTGAT |
7 | TCGCCTGATAAATTGTGTCGAAATCC TTCTCTTGGAAAGAAAGTGAT |
SEQIDNO: | 表2捕获扩展物库B |
8 | TCCAAAAAAAAAGGCTCCAAAAGGAGCCTTTATTCTCTTGGAAAGAAAGTGAT |
9 | CGCCGACAATGACAACAACCATCGCTTCTCTTGGAAAGAAAGTGAT |
SEQIDNO: | 表3标记扩展物库 |
10 | ATGAGGAAGTTTCCATTAAACGGGTTTAGGCATAGGACCCGTGTCT |
11 | GAGGCTTTGAGGACTAAAGACTTTTTC TTAGGCATAGGACCCGTGTCT |
12 | CCCAGCGATTATACCAAGCGCG TTAGGCATAGGACCCGTGTCT |
13 | AAGAATACACTAAAACACTCATCTTTGACCTTAGGCATAGGACCCGTGTCT |
14 | CTTTGAAAGAGGACAGATGAACGGTG TTAGGCATAGGACCCGTGTCT |
15 | GGAACGAGGCGCAGACGGTCA TTAGGCATAGGACCCGTGTCT |
16 | ACGAGGGTAGCAACGGCTACA TTAGGCATAGGACCCGTGTCT |
17 | GCGACCTGCTCCATGTTACTTAGCCTTAGGCATAGGACCCGTGTCT |
18 | CTCAGCAGCGAAAGACAGCATCGGATTAGGCATAGGACCCGTGTCT |
19 | ATCATAAGGGAACCGAACTGACCAATTAGGCATAGGACCCGTGTCT |
20 | CCACGCATAACCGATATATTCGGTCTTAGGCATAGGACCCGTGTCT |
21 | TACAGACCAGGCGCATAGGCTGGCTTAGGCATAGGACCCGTGTCT |
22 | AAACAAAGTACAACGGAGATTTGTATCATTAGGCATAGGACCCGTGTCT |
23 | CACCAACCTAAAACGAAAGAGGCGATTAGGCATAGGACCCGTGTCT |
24 | AAAATACGTAATGCCACTACGAAGGTTAGGCATAGGACCCGTGTCT |
表4非专一性结合试验背景噪声 | ||
捕获扩展物库 | 信号(+M13噬菌体) | 噪声(-M13噬菌体) |
只有Pool A | 293,306,337,359 | 1.1,0.9,1.1,2.0 |
Pool A+Pool B | 390,393,379,376 | 103,130,436,172 |
表4iosC*isoG碱基配对专一性的Tm分析 | |||
匹配/错配,成对寡聚核苷酸 | Tm1 | Tm2 | 平均Tm |
C*G 匹配,1*3 | 60 | 60 | 60 |
isoC*isoG 匹配,2*4 | 60 | 61 | 60 |
isoC*G 错配,2*3 | 52 | 52 | 52 |
isoG*C 错配,1*4 | 52 | 52 | 52 |
G*T 错配,3*5 | 50 | 49 | 49 |
isoG*T 错配,4*5 | 53 | 53 | 53 |
表5isoC*isoG碱基配对的pH敏感性的Tm分析 | |||||
杂交物,成对的寡聚核苷酸 | pH | Tm1 | Tm2 | Tm3 | 平均Tm |
isoG*isoC,2*4 | 7.9 | 62 | 60 | 60 | 61 |
isoG*isoC,2*4 | 5.1 | 60 | 59 | 62 | 60 |
isoG*isoC,2*4 | 9.5 | 53 | 51 | 52 | 52 |
G*C,1*3 | 9.5 | 52 | 52 | 52 |
表6作为盐浓度函数的isoC*isoG的稳定性 | ||||
杂交物,成对寡聚核苷酸 | 盐(M) | pH | 平均Tm(℃) | dTmdlog[Na+] |
isoC*isoG,2*4 | 0.5 | 7.9 | 61 | |
isoC*isoG,2*4 | 0.17 | 7.9 | 56 | 10-11 |
isoC*isoG,2*4 | 0.5 | 9.5 | 52 | |
isoC*isoG,2*4 | 0.17 | 9.5 | 50 | 3 |
C*G,1*3 | 0.5 | 9.5 | 52 | |
C*G,1*3 | 0.1 | 9.5 | 48.5 | 5 |
表7 | |
SEQIDNO: | 序列1 |
30 | 5′ GATGTGGTTGTCGTACTTTTTTTGACACTCCACCAT |
31 | 5′ GATGTGGTTGTCGTACTTTTTTTGACAFTCCJCCAT |
32 | ALK.PHOS.-CTACACCAACAGCATGAA 5′ |
33 | 3′TCACTAAGTACCACCTCACAG |
34 | 5′AGTGATTCATGGTGGAGTGTCTCTCTTGGAAAGAAAGTGAT |
35 | 3′ GAGAACCTTTCTTTCACTX |
表8碱基对错配对杂交的影响 | ||||
样本号NO: | 初次杂交 | 二次杂交 | 平均RLU1(n=6) | %CV2 |
1 | 34+35 | 30+32 | 399 | 7 |
2 | 33+34+35 | 30+32 | 30 | 9 |
3 | 34+35 | 31+32 | 9 | 6 |
4 | 33+34+35 | 31+32 | 0.4 | 4 |
5 | 34+35 | 32 | 0.1 | 11 |
序列(5′-->3′) | 序列ID |
AGT FAJ CGC FGT AFC AAJTJC.......... | AMP重复序列(臂) |
ATC ACG AAC TCA TCA CGA ACT C | AMP引导序列(梳) |
GFA FTT GJT ACJ GCG FTJ ACT......L | AP探针序列 |
试验 | 当前检测极限 | IsoC/G检测极限 | 提高倍数 |
IL-2 | 41,000 | 3,000 | 12.6 |
IL-6 | 34,000 | 11,000 | 3.0 |
Claims (13)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/298,073 | 1994-08-30 | ||
US08/298,073 US5681702A (en) | 1994-08-30 | 1994-08-30 | Reduction of nonspecific hybridization by using novel base-pairing schemes |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1164260A true CN1164260A (zh) | 1997-11-05 |
CN100335649C CN100335649C (zh) | 2007-09-05 |
Family
ID=23148897
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNB951958801A Expired - Lifetime CN100335649C (zh) | 1994-08-30 | 1995-08-30 | 在核酸杂交试验中减少非特异性杂交 |
Country Status (21)
Country | Link |
---|---|
US (4) | US5681702A (zh) |
EP (2) | EP0778898B1 (zh) |
JP (4) | JP4461278B2 (zh) |
KR (1) | KR100218113B1 (zh) |
CN (1) | CN100335649C (zh) |
AT (2) | ATE259374T1 (zh) |
AU (1) | AU708194B2 (zh) |
BG (1) | BG101246A (zh) |
BR (1) | BR9508674A (zh) |
CA (1) | CA2197901A1 (zh) |
CZ (1) | CZ58997A3 (zh) |
DE (2) | DE69528839T2 (zh) |
ES (2) | ES2187571T3 (zh) |
FI (1) | FI970803A (zh) |
HU (1) | HUT77754A (zh) |
MX (1) | MX9701419A (zh) |
NO (1) | NO970884L (zh) |
NZ (1) | NZ292451A (zh) |
PL (1) | PL318933A1 (zh) |
SK (1) | SK25497A3 (zh) |
WO (1) | WO1996006950A1 (zh) |
Cited By (4)
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CN104053777A (zh) * | 2011-11-18 | 2014-09-17 | 塔古西库斯生物株式会社 | 与靶蛋白质结合的核酸片段 |
CN110577981A (zh) * | 2018-06-07 | 2019-12-17 | 北京福安华生物科技有限公司 | 一种基于人工模拟核酸的分子信标 |
CN111378746A (zh) * | 2018-12-27 | 2020-07-07 | 北京福安华生物科技有限公司 | 检测CASP7基因rs12415607位点多态性的人工模拟核酸分子信标与试剂盒 |
WO2023246124A1 (zh) * | 2022-06-23 | 2023-12-28 | 南京浦光生物科技有限公司 | 检测目标抗体的试剂组合、试剂盒、检测系统及检测方法 |
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