title: Single Lysis-Salting Out Method of Genomic DNA Extraction From Dried Blood Spots.
abstract: BACKGROUND
  Dried blood spots (DBS) are an important form of bio-sampling and valuable approach for storing blood samples for genetic studies. This has necessitated in developing an effective protocol to isolate genomic DNA (gDNA) from DBS samples.In this study, we have elucidated a dependable and non-hazardous "single lysis-salting out" (SLSO) protocol of gDNA extraction from DBS and compared against the available commercial kits.
  
  
  METHODS
  For the purpose of this study, blood spots were collected on S&S 903 filter cards from 10 healthy volunteers and 30 patients with glutaric aciduria type I (GA-I). The gDNA was extracted from theseDBS samples by SLSO, QIAamp® gDNA Micro kit and innuPREP forensic kit methods. The quantity and quality of gDNA obtained from these methods were determined by measuring the absorbance using a Nanodrop spectrophotometer.
  
  
  RESULTS
  The SLSO method showed four-fold and eight-fold increased yield of gDNA in healthy volunteers and patient samples, respectively, compared to commercial kits (p<0.0001). The protocol was also found to be cost efficient, reducing the per sample cost to almost half. The suitability of this method for genetic studies was confirmed by performing R402W genotyping by RFLP in GA-I patients. The genotyping results showed the presence of R402W mutation in 20% (6/30) of patients.
  
  
  CONCLUSIONS
  The SLSO method was found to be inexpensive, non-hazardous and a suitable technique for isolating gDNA from DBS samples for genetic studies.
authors.affiliation: Department of Neurochemistry, National Institute of Mental Health and Neurosciences, Bengaluru, Karnataka, India.
authors.forename: Muntaj
authors.initials: M
authors.lastname: Shaik
authors.affiliation: Department of Biochemistry, Karnatak University Dharwad, Dharwad, Karnataka, India.
authors.forename: Devaraju Kuramkote
authors.initials: DK
authors.lastname: Shivanna
authors.affiliation: Department of Pediatrics, Jawaharlal Nehru Medical College, KLE University, Belgaum, Karnataka, India.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Kamate
authors.affiliation: Department of Biotechnology & Microbiology, Karnatak University Dharwad, Dharwad, Karnataka, India.
authors.forename: Vedamurthy
authors.initials: V
authors.lastname: Ab
authors.affiliation: Department of Neurochemistry, National Institute of Mental Health and Neurosciences, Bengaluru, Karnataka, India. kruthikav-tp@nimhans.ac.in, kruthikavinod@gmail.com.
authors.forename: Kruthika-Vinod
authors.initials: KV
authors.lastname: Tp
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000097:blood* / Q000235:genetics*; D000704:Analysis of Variance; D001928:Brain Diseases, Metabolic / Q000097:blood* / Q000235:genetics*; D004247:DNA / Q000097:blood*; D059788:Dried Blood Spot Testing / Q000379:methods*; D005260:Female; D023281:Genomics / Q000379:methods*; D005838:Genotype; D050770:Glutaryl-CoA Dehydrogenase / Q000097:blood / Q000172:deficiency* / Q000235:genetics; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D009154:Mutation / Q000235:genetics*; D013048:Specimen Handling
chemical_list: D004247:DNA; D050770:Glutaryl-CoA Dehydrogenase
keywords: DNA isolation; GCDH; R402W mutation; genotyping; guthrie cards