Nucleic Acids Isolation and DNA Analysis Facility

University Of Tx Md Anderson Can Ctr, Houston TX

Investigators

Linked publications & trials

Abstract

[unreadable] The specific goals for Core C are:[unreadable] 1) establishment of lymphoblastoid cell lines and fibroblast cultures on LFS and WT patients.[unreadable] 2) isolation of DNA, RNA, and plasma from peripheral blood; and DNA and RNA from tumor and normal[unreadable] tissue samples from study individuals. This includes those with Wilms tumors, those who are[unreadable] potential p53 mutation carriers, and family members of these individuals.[unreadable] 3) mutation detection by direct sequencing of PCR products for p53, WT1, and/or some WT "pathway"[unreadable] genes for childhood sarcoma kindreds, Wilms tumor kindreds, and sporadic Wilms tumors[unreadable] 4) Southern analysis of WT patients to identify WT1 deletions undetectable by PCR-based sequencing.[unreadable] 5) functional characterization of odd p53 missense mutations and identification of potentially truncated[unreadable] proteins in cells from individuals carrying nonsense and splicing mutants[unreadable] 6) mutation screening by WAVE analysis for selected WT pathway genes identified in Project 4[unreadable] 7) storage and distribution, as needed, of DNA, RNA, and plasma samples[unreadable] 8) maintenance of a database detailing sample handling, nucleic acids isolations, and mutational[unreadable] analysis. This database will be used to update the Program's database managed by Core C.[unreadable] 9) archiving of sequence and mutation screening output.[unreadable] These functions are critical to the success of all projects within the P01. Project 1 uses p53 and WT1 data[unreadable] to characterize better cancer risk and also to develop better genetic models for cancer predisposition. The[unreadable] information regarding p53 mutational status will also identify non-p53 families that will aid the identification[unreadable] (Proj. 2) of other genes responsible for predisposition and guide the interpretation of the tumor data[unreadable] generated from mutant mice (Projects 3 & 5). WT1 mutational status is critical for the delineation of the[unreadable] molecular subsets of Wilms tumors and WT patients and elucidating the biologic effect of the pattern of[unreadable] mutations, alterations, and expression changes we have previously identified in Wilms tumors (Project 4).[unreadable] This core provides sample processing, mutation screening and analyses, functional analyses, sample[unreadable] storage, and data storage critical for all projects of the P01.