Evaluation of quality of DNA extracted from buccal swabs

Page 1

Scientific Publication Suitability of INFOHaem using blood DNA instead of buccal cell DNA

Evaluation of quality of DNA extracted from buccal swabs for microarray based genotyping Livy Alex et al. (2011) Ind J Clin Biochem 27: 28-33

Full article published in:

Indian Journal of Clinical Biochemistry Buccal cell collection as a source of DNA was initially put forward as an efficient means of costeffective DNA collection. However INFOHaem Molecular Screening Services prefer blood DNA to be collected for genetic predisposition and diagnostic tests. Although non-invasive DNA collection methods are preferred over phlebotomy, there have been conflicting reports about the methodology and results obtained from using buccal DNA. It is not very clear if phlebotomy can be confidently replaced by buccal cell DNA. To address this issue, this study compared the performance of buccal and blood DNA from same subjects in a genotyping experiment and this paper reports the results.

Subject ID

Buccal

Cotton swab derived buccal cells were scraped from the inner side of cheeks from 16 subjects, and blood was also drawn from the same 16 subjects participating in a genotypic association study of a lipid disease. The DNA quality was assessed by resolving on agarose gels, checking purity (A260/A280) and finally by microarray hybridization. This study, by INFOVALLEY® , recommends increasing the total amount of DNA used as starting material in case of buccal DNA, at least greater than 250 ng. However, blood has the advantage that the yield of DNA is better and enough of reserve samples is available to repeat the experiment which is not possible with buccal samples. Genotyping results can be seriously compromised if care is not taken to check the quality and yields of such specimens. Therefore, blood is a better alternative than buccal DNA.

Blood

Concentration (ng/μl)

Purity (A260/280)

Concentration (ng/μl)

Purity (A260/280)

1

257.96

1.28

51.26

2.0

2

296.86

1.22

30.36

2.1

3

172.6

1.3

100.76

1.9

4

26.03

0.9

98.03

1.6

5

166.8

1.3

91.12

2.0

6

106.23

1.3

61.3

1.5

7

433.2

1.9

78

1.7

8

166.8

1.8

51.4

2.0

9

185.03

1.4

100.86

1.5

10

129.16

2.0

100.66

1.7

11

63.33

1.2

100.96

1.6

12

98.36

1.2

87.6

1.5

13

33.6

1.1

67.8

1.6

14

18.95

0.8

93.05

1.7

15

58.6

1.2

76.16

1.6

16

29.8

1.3

65.9

1.6

The purity of the buccal DNA is less that 1.7, with an average reading of 1.3, which indicates that there is protein contamination. However, blood DNA purity seems to be good averaging at 1.7

Comparison of buccal and blood DNA yields and quality. © 2012 INFOVALLEY. www.infovalley.net.my


Turn static files into dynamic content formats.

Create a flipbook
Issuu converts static files into: digital portfolios, online yearbooks, online catalogs, digital photo albums and more. Sign up and create your flipbook.