OBJECTIVES: The objectives of this paper are to determine the storage stability of saliva at 37 °C over an 18-month period, and its influence on the DNA yield, purity, PCR protocols and genotyping efficacy. MATERIALS AND METHODS: Of the 60 participants, blood samples were obtained from 10 and saliva from 50. Samples were subjected to different storage conditions: DNA extracted immediately; DNA extracted following storage at 37 °C for 1, 6, 12 and 18 months. Subsequently, DNA yield, OD(260/280) and OD(260/230) ratios were measured. The isolated DNA was used to amplify exons 0-7 of the RUNX2 gene and subsequently sequenced. Furthermore, 25 SNPs were genotyped. RESULTS: The mean DNA yield, OD(260/280) and OD(260/230) ratios obtained from blood were 67.4 ng/μl, 1.8 ± 0.05 and 1.8 ± 0.4 respectively. DNA yield obtained from saliva was significantly higher than blood (p < 0.0001), ranging from 97.4 to 125.8 ng/μl while the OD(260/280) ratio ranged from 1.8 ± 0.13 to 1.9 ± 0.1. The success rates for the 25 SNPs ranged from 98 to 100 % for blood and 96-99 % for saliva samples with the genotype frequencies in Hardy-Weinberg equilibrium (>0.01). CONCLUSIONS: Saliva can be stored at 37 °C for 18 months without compromising its quality and ability to endure genetic analyses. CLINICAL RELEVANCE: Saliva is a viable source of human DNA to facilitate the feasibility of large-scale genetic studies.
OBJECTIVES: The objectives of this paper are to determine the storage stability of saliva at 37 °C over an 18-month period, and its influence on the DNA yield, purity, PCR protocols and genotyping efficacy. MATERIALS AND METHODS: Of the 60 participants, blood samples were obtained from 10 and saliva from 50. Samples were subjected to different storage conditions: DNA extracted immediately; DNA extracted following storage at 37 °C for 1, 6, 12 and 18 months. Subsequently, DNA yield, OD(260/280) and OD(260/230) ratios were measured. The isolated DNA was used to amplify exons 0-7 of the RUNX2 gene and subsequently sequenced. Furthermore, 25 SNPs were genotyped. RESULTS: The mean DNA yield, OD(260/280) and OD(260/230) ratios obtained from blood were 67.4 ng/μl, 1.8 ± 0.05 and 1.8 ± 0.4 respectively. DNA yield obtained from saliva was significantly higher than blood (p < 0.0001), ranging from 97.4 to 125.8 ng/μl while the OD(260/280) ratio ranged from 1.8 ± 0.13 to 1.9 ± 0.1. The success rates for the 25 SNPs ranged from 98 to 100 % for blood and 96-99 % for saliva samples with the genotype frequencies in Hardy-Weinberg equilibrium (>0.01). CONCLUSIONS: Saliva can be stored at 37 °C for 18 months without compromising its quality and ability to endure genetic analyses. CLINICAL RELEVANCE: Saliva is a viable source of human DNA to facilitate the feasibility of large-scale genetic studies.
Authors: M S Phillips; R Lawrence; R Sachidanandam; A P Morris; D J Balding; M A Donaldson; J F Studebaker; W M Ankener; S V Alfisi; F-S Kuo; A L Camisa; V Pazorov; K E Scott; B J Carey; J Faith; G Katari; H A Bhatti; J M Cyr; V Derohannessian; C Elosua; A M Forman; N M Grecco; C R Hock; J M Kuebler; J A Lathrop; M A Mockler; E P Nachtman; S L Restine; S A Varde; M J Hozza; C A Gelfand; J Broxholme; G R Abecasis; M T Boyce-Jacino; L R Cardon Journal: Nat Genet Date: 2003-02-18 Impact factor: 38.330
Authors: M B Hocaoglu; S Gurkas; T Karaderi; B Taneri; K Erguler; B Barin; E M Bilgin; G Eralp; M Allison; N Findikli; K Boynukalin; M Bahceci; H Naci; K Vincent; S A Missmer; C M Becker; K T Zondervan; N Rahmioglu Journal: BMC Womens Health Date: 2019-04-03 Impact factor: 2.809