Unknown

Dataset Information

0

The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when?


ABSTRACT: Museums and pathology collections around the world represent an archive of genetic material to study populations and diseases. For preservation purposes, a large portion of these collections has been fixed in formalin-containing solutions, a treatment that results in cross-linking of biomolecules. Cross-linking not only complicates isolation of nucleic acid but also introduces polymerase "blocks" during PCR. A wide variety of methods exists for the recovery of DNA and RNA from archival tissues, and although a number of previous studies have qualitatively compared the relative merits of the different techniques, very few have undertaken wide scale quantitative comparisons. To help address this issue, we have undertaken a study that investigates the quality of nucleic acids recovered from a test panel of fixed specimens that have been manipulated following a number of the published protocols. These include methods of pre-treating the samples prior to extraction, extraction and nucleic acid purification methods themselves, and a post-extraction enzymatic repair technique. We find that although many of the published methods have distinct positive effects on some characteristics of the nucleic acids, the benefits often come at a cost. In addition, a number of the previously published techniques appear to have no effect at all. Our findings recommend that the extraction methodology adopted should be chosen carefully. Here we provide a quick reference table that can be used to determine appropriate protocols for particular aims.

SUBMITTER: Gilbert MT 

PROVIDER: S-EPMC1888728 | biostudies-literature | 2007 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when?

Gilbert M Thomas P MT   Haselkorn Tamara T   Bunce Michael M   Sanchez Juan J JJ   Lucas Sebastian B SB   Jewell Laurence D LD   Van Marck Eric E   Worobey Michael M  

PloS one 20070620 6


Museums and pathology collections around the world represent an archive of genetic material to study populations and diseases. For preservation purposes, a large portion of these collections has been fixed in formalin-containing solutions, a treatment that results in cross-linking of biomolecules. Cross-linking not only complicates isolation of nucleic acid but also introduces polymerase "blocks" during PCR. A wide variety of methods exists for the recovery of DNA and RNA from archival tissues,  ...[more]

Similar Datasets

| S-EPMC6128582 | biostudies-literature
| S-EPMC6801416 | biostudies-literature
| S-EPMC7119673 | biostudies-literature
| S-EPMC5552883 | biostudies-literature
| S-EPMC7438656 | biostudies-literature
| S-EPMC2996486 | biostudies-literature
| S-EPMC2329784 | biostudies-literature
| S-EPMC7236785 | biostudies-literature
| S-EPMC4778669 | biostudies-literature
| S-EPMC2963916 | biostudies-literature