FULL LENGTH CDNA SEQUENCING FOR PEDIATRIC LEUKEMIAS

儿童白血病的全长 CDNA 测序

• 批准号: 6173771
• 负责人: RICHARD A GIBBS
• 金额: $ 140.79万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173771
• 关键词: clinical research; complementary DNA; genetic library; human genetic material tag; human subject; leukemia; messenger RNA; neoplasm /cancer genetics; nucleic acid sequence; pediatric neoplasm /cancer

DESCRIPTION: (Applicant's Description) This project aims to generate cDNA libraries from fresh pediatric leukemia cells that have been directly isolated from blood at the time of diagnosis. Cases that represent the major subtypes of childhood Acute Lymphoid leukemia (ALL) and at least one subtype of Acute Myelogenous Leukemia (AML) will be chosen for analysis. The library construction methodology will utilize improvements on existing techniques which have been demonstrated to generate libraries with a high proportion of foil-length clones. Clones from these libraries will contain full-length copies of the original mRNA, and the libraries will be representative of the original messenger population. The libraries will be normalized to reduce the variation in abundance of different clones and aliquots from non-normalized and normalized libraries will be arrayed to facilitate replication and identification of clones of interest. EST sequencing on 10,000 randomly selected clones will be performed each year to select cDNAs that appear to be full- length, and for which there is no existing foil length sequence available in the databases. The full length sequence of >350 1.5-2.0 kb cDNAs will be determined each year, using a concatenation cDNA sequencing approach that was developed in this laboratory. This protocol enables us to simultaneously perform full-length sequencing on as many as 70 cDNAs after construction of a single shotgun library. A PCR-based method that has been optimized m our laboratory will be used to identify the 5'-ends of clones that are not full length. All data will be released immediately and all clones will be freely available. These malignant tissues have not been extensively sequenced, and can therefore be expected to provide substantial new information compared to that derived from leukemia cell lines. Hence, the availability of these libraries will provide a mechanism for rapidly isolating full-length copies of previously identified partial clones, and will provide a source for identifying new cancer-related genes. The integration of the tissue collection, library construction, EST analysis, full length clone insert sequencing and 5'- end rescue will define a complete and scalable pathway for the systematic analysis of genes involved in cancer.

描述：（申请人描述）本项目旨在生成cDNA

项目成果

LNCIB human full-length cDNAs collection: towards a better comprehension of the human transcriptome.

LNCIB 人类全长 cDNA 集合：更好地理解人类转录组。

• DOI: 10.1016/j.crvi.2003.09.029
• 发表时间: 2003
• 期刊: Comptes rendus biologies
• 影响因子: 2
• 作者: Dalla,Emiliano;Verardo,Roberto;Lazarević,Dejan;Marchionni,Luigi;Reid,JamesF;Bahar,Nabil;Klarić,Enio;Marcuzzi,Giacomo;Marzio,Riccardo;Belgrano,Anna;Licastro,Danilo;Schneider,Claudio
• 通讯作者: Schneider,Claudio

Concatenation cDNA sequencing for transcriptome analysis.

用于转录组分析的串联 cDNA 测序。

• DOI: 10.1016/j.crvi.2003.09.032
• 发表时间: 2003
• 期刊: Comptes rendus biologies
• 影响因子: 2
• 作者: Gunaratne,PreethiH;Wu,JiaQian;Garcia,AngelaM;Hulyk,Steven;Worley,KimC;Margolin,JudithF;Gibbs,RichardA
• 通讯作者: Gibbs,RichardA