New Jersey Center for Tourette Syndrome Sharing Repository: Methods and Sample Description
Heiman GA, King RA, Tischfield JA.
Department of Genetics, Rutgers University, 145 Bevier Road, Piscataway, NJ 08854, USA. heiman@biology.rutgers.edu
Abstract
BACKGROUND:
Tourette Syndrome is a neuropsychiatric disorder characterized by chronic motor and phonic tics. Affected individuals and their family members are at an increased risk for other neuropsychiatric conditions including obsessive-compulsive disorder and attention deficit hyperactivity disorder. While there is consistent evidence that genetic factors play a significant etiologic role, no replicable susceptibility alleles have thus far been identified.
DESCRIPTION:
Here we discuss a sharing resource of clinical and genetic data, the New Jersey Center for Tourette Syndrome Sharing Repository, whose goal is to provide clinical data, DNA, and lymphoblastoid cell lines to qualified researchers.
CONCLUSION:
Opening access to the data and patient material to the widest possible research community will hasten the identification of causal genetic factors and facilitate better understanding and treatment of this often impairing disorder.
http://www.ncbi.nlm.nih.gov/pubmed/19036136
Preparing DNA from Mammalian Sources for Genotyping
Sahota A, Brooks AI, Tischfield JA.
Rutgers University Cell and DNA Repository, Department of Genetics, Piscataway, NJ 08854-8082, USA.
Abstract
INTRODUCTION
The availability of high-quality DNA from a large number of individuals is a prerequisite for the success of genetic variation studies. This requirement has spurred major technological advances in DNA extraction methodologies. Twenty years ago, large-scale manual extractions took >3 d to complete. Large-scale preparations can now be completed within 3 h using automated extraction platforms, without the need for toxic reagents and often with higher yields than in the past. This article introduces methods and considerations for the extraction of DNA for genotyping, and for the determination of DNA quantity and quality.
http://www.ncbi.nlm.nih.gov/pubmed/21357154
Preparing DNA From Saliva for Genotyping
Sahota A, Brooks AI, Tischfield JA.
Rutgers University Cell and DNA Repository, Department of Genetics, Piscataway, NJ 08854-8082, USA.
Abstract
INTRODUCTION
Saliva may be a viable alternative to blood as a source for DNA, and has the advantage that it is collected noninvasively. This protocol describes the Oragene procedure for DNA extraction from 2-mL saliva samples. It yields high-quality DNA suitable for research and diagnostic applications.
http://www.ncbi.nlm.nih.gov/pubmed/21357152
Preparing DNA From Blood for Genotyping
Sahota A, Brooks AI, Tischfield JA, King IB.
Rutgers University Cell and DNA Repository, Department of Genetics, Piscataway, NJ 08854-8082, USA.
Abstract
INTRODUCTION
This protocol describes the extraction of genomic DNA from whole blood samples (fresh or frozen) and buffy coats. It provides methods for (1) large-scale extraction of DNA using either in-house or commercial (PUREGENE) reagents; (2) mid-scale extraction of DNA using the QIAamp DNA Blood Midi Kit (for 0.5-mL samples); and (3) small-scale extraction of DNA using the QIAamp DNA Blood Mini Kits (for 200-μL samples). Also included are methods for extracting DNA from blood samples that are compromised or clotted. Compromised blood samples include samples that spent more than 3 days in transit, samples that have been stored for more than 1 wk at 4°C, samples that have been stored at -20°C or -70°C, and samples that were collected incorrectly (e.g., not mixed properly). The procedures described here are not high throughput. Potential drawbacks of making them high-throughput are lower recoveries of DNA and possible variability in data quality, thus requiring additional quality assurance and control checks. DNA yields for these methods vary with the number of nucleated cells in the blood sample and the quality of the sample.
http://www.ncbi.nlm.nih.gov/pubmed/21357151
Preparing DNA from Cell Pellets for Genotyping
Sahota A, Brooks AI, Tischfield JA. Rutgers University Cell and DNA Repository, Department of Genetics, Piscataway, NJ 08854-8082, USA.
Abstract
INTRODUCTION
This protocol describes large-scale DNA extraction from cell pellets using either in-house or commercial (PUREGENE) reagents. The cell pellets are prepared from human lymphoblast cell lines (LCLs) that have been established using the Epstein-Barr virus (EBV).
http://www.ncbi.nlm.nih.gov/pubmed/21357150
