Tissue preparation for laser capture microdissection and RNA extraction from fresh frozen breast tissue

Tissue preparation for laser capture microdissection and RNA extraction from fresh frozen breast tissue.

A robust RNA integrity-preserving staining protocol for laser capture microdissection of endometrial cancer tissue.

Laser capture microdissection of frozen tissue sections allows homogeneous cell populations to be isolated for expression profiling. However, this requires striking a balance between retaining adequate morphology for accurate microdissection and maintaining RNA integrity. Various staining protocols were applied to frozen endometrial carcinoma tissue sections. Although alcohol-based methods were...

Optimization of RNA Extraction from Rat Pancreatic Tissue

Background: Optimized RNA extraction from tissues and cell lines consists of four main stages regardless of the method of extraction: 1) homogenizing, 2) effective denaturation of proteins from RNA, 3) inactivation of ribonuclease, and 4) removal of any DNA, protein, and carbohydrate contamination. Isolation of undamaged intact RNA is challenging when the related tissue contains high levels of ...

Differential Gene Expression Analysis of Pure Cell Populations from Frozen Uterine Tissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays

Laser Capture Microdissection (LCM) is an innovative technique which permits the rapid and reliable procurement of pure cell populations from heterogeneous tissue sections. After LCM, RNA isolated from pure cell populations was amplified to generate adequate amounts of antisense RNA (aRNA) for hybridization, and the differential gene expression profiles were analyzed between two experimental gr...

Cytoplasmic RNA extraction from fresh and frozen mammalian tissues.

The quality of collections of expressed sequence tags andfull-length cDNAs is adversely affected by the presence of "junk" clones derivedfrom unspliced or partially spliced RNAs present in conventional total RNA preparations. One can overcome this problem by using intact cytoplasmic RNA to create cDNA libraries, but the methods in the literature that describe the preparation of RNA only work we...