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1. (WO2019061196) PCR PRIMERS FOR DETECTION AND USE THEREOF
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Pub. No.: WO/2019/061196 International Application No.: PCT/CN2017/104105
Publication Date: 04.04.2019 International Filing Date: 28.09.2017
IPC:
C12N 15/11 (2006.01) ,C12Q 1/68 (2018.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
68
involving nucleic acids
Applicants:
深圳华大生命科学研究院 BGI SHENZHEN [CN/CN]; 中国广东省深圳市 盐田区北山工业区综合楼 Main Building, Beishan Industrial Zone, Yantian District Shenzhen, Guangdong 518083, CN
Inventors:
周清 ZHOU, Qing; CN
王飞 WANG, Fei; CN
王磊 WANG, Lei; CN
尹悦露 YIN, Yuelu; CN
吴靓 WU, Liang; CN
赵正琦 CHAO, Cheng-chi; CN
李贵波 LI, Guibo; CN
杨乃波 YANG, Naibo; CN
侯勇 HOU, Yong; CN
李波 LI, Bo; CN
Agent:
北京品源专利代理有限公司 BEYOND ATTORNEYS AT LAW; 中国北京市 海淀区莲花池东路39号西金大厦6层 F6, Xijin Centre 39 Lianhuachi East Rd. , Haidian District Beijing 100036, CN
Priority Data:
Title (EN) PCR PRIMERS FOR DETECTION AND USE THEREOF
(FR) AMORCES DE PCR POUR LA DÉTECTION ET LEUR UTILISATION
(ZH) 一种用于检测的PCR引物及其应用
Abstract:
(EN) Disclosed are PCR primers for detection and a use thereof, which primers comprise an upstream primer and a downstream primer; the upstream primer is a primer designed according to a linker sequence which is introduced into the 3' end of a first chain of cDNA; and the downstream primer is a downstream primer sequence designed according to the 3' end of a target fragment to be detected. The primers and method of the present invention are effective for different target fragments, do not require specific upstream primers to be designed for single different target fragments, and introduce a fixed IS sequence into the 3' end of a transcript first chain of cDNA of an mRNA chain by using SMART template conversion technology during a reverse transcription process, and subsequent PCR uses a complementary sequence of the IS sequence as the downstream primer while cooperating with the specific downstream primer, successfully amplifying the target fragment.
(FR) L'invention concerne des amorces de PCR pour la détection et une utilisation de celles-ci, lesquelles amorces comprennent une amorce amont et une amorce aval ; l'amorce amont étant une amorce conçue selon une séquence de lieur qui est introduite dans l'extrémité 3' d'une première chaîne d'ADNc ; et l'amorce aval étant une séquence d'amorce aval conçue selon l'extrémité 3' d'un fragment cible à détecter. Les amorces et le procédé selon la présente invention sont efficaces pour différents fragments cibles, ne nécessitent pas de concevoir des amorces amont spécifiques pour différents fragments cibles individuels et introduisent une séquence IS fixe à l'extrémité 3' d'une première chaîne de transcrit d'ADNc d'une chaîne d'ARNm à l'aide de la technologie de conversion de matrice SMART pendant un processus de transcription inverse, et la PCR ultérieure utilise une séquence complémentaire de la séquence IS en tant qu'amorce aval tout en coopérant avec l'amorce aval spécifique, ce qui permet d'amplifier avec succès le fragment cible.
(ZH) 本发明公开了一种用于检测的PCR引物及其应用,包括上游引物和下游引物;所述上游引物为根据一段接头序列设计的引物,所述接头序列引入到第一链cDNA的3'端;所述下游引物为根据检测的目的片段的3'端设计的下游引物序列。本发明引物和方法对不同目的片段均有效,不需要针对单一不同目的片段设计特异性的上游引物,通过在反转录过程中,采用SMART模板转换技术使得mRNA链的转录本第一链cDNA的3'端引入一段固定IS序列,后续PCR均以IS序列的互补序列为上游引物,同时配合特异性的下游引物,成功扩增出目的片段。
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Chinese (ZH)
Filing Language: Chinese (ZH)