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Sazonova et al. Vessel Plus 2019;3:5 I http://dx.doi.org/10.20517/2574-1209.2018.56 Page 3 of 9

Clinical, anthropometric and age characteristics were determined for patients with left ventricular
hypertrophy and conventionally healthy individuals [Tables 1 and 2]

The study was carried out in accordance with the Declaration of Helsinki. The study protocol was inspected
and approved by the Ethics Committee of the National Medical Research Center of Cardiology. Each study
participant has signed a written informed consent to participate in this investigation.

DNA from white blood cells was isolated using the phenol-chloroform method [32-34] . PCR-fragments of
DNA were obtained. They contained the region of studied mutations (652insG, m.5178C>A, m.3336T>C,
m.14459G>A, 652delG, m.14846G>A, m.1555A>G, m.15059G>A, m.3256C>T, m.12315G>A and m.13513G>A).
It should be noted that biotin was attached to one of the DNA chains of the PCR fragment using the primer
“bio-” (biotinylated). This was necessary for the analysis of the biotinylated DNA chain of the investigated
amplificate using the pyrosequencing [35-37] .

PCR primer sequences, taken for the present reseach [24,26,28-30] :
1. For m.652insG
F: TAGACGGGCTCACATCAC (621-638)
R: bio-GGGGTATCTAATCCCAGTTTGGGT (1087-1064)
2. For m.5178C>A
F: bio-GCAGTTGAGGTGGATTAAAC (4963-4982)
R: GGAGTAGATTAGGCGTAGGTAG (5366-5345)
3. For m.3336T>C
F: bio-AGGACAAGAGAAATAAGGCC (3129-3149)
R: ACGTTGGGGCCTTTGCGTAG (3422-3403)
4. For m.14459G>A
F: CAGCTTCCTACACTATTAAAGT (14303-14334)
R: bio-GTTTTTTTAATTTATTTAGGGGG (14511-14489)
5. For m.652delG
F: TAGACGGGCTCACATCAC (621-638)
R: bio-GGGGTATCTAATCCCAGTTTGGGT (1087-1064)
6. For m.14846G>A
F: bio-CATTATTCTCGCACGGACT (14671-14689)
R: GCTATAGTTGCAAGCAGGAG (15120-15100)
7. For m.1555A>G
F: TAGGTCAAGGTGTAGCCCATGAGGTGGCAA (1326-1355)
R: bio-GTAAGGTGGAGTGGGTTTGGG (1704-1684)
8. For m.15059G>A
F: bio-CATTATTCTCGCACGGACT (14671-14689)
R: GCTATAGTTGCAAGCAGGAG (15120-15100)
9. For m.3256C>T
F: bio-AGGACAAGAGAAATAAGGCC (3129-3149)
R: ACGTTGGGGCCTTTGCGTAG (3422-3403)
10. For m.12315G>A
F: bio-CTCATGCCCCCATGTCTAA (12230-12249)
R: TTACTTTTATTTGGAGTTGCAC (12337-12317)
11. For m.13513G>A
F: CCTCACAGGTTTCTACTCCAAA (13491-13512)
R: bio-AAGTCCTAGGAAAGTGACAGCGAGG (13825-13806)