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Investigation of optimum concentrations of betaine for improving the resolution of sequencing G-C rich DNA with trinucleotide repeats / 中华医学遗传学杂志
Chinese Journal of Medical Genetics ; (6): 163-169, 2014.
Article in Chinese | WPRIM | ID: wpr-254490
ABSTRACT
<p><b>OBJECTIVE</b>To develop an optimal sequencing system which can improve the resolution of sequencing G-C rich DNA with abundant trinucleotide repeats by applying concentration gradients of betaine to the Sanger sequencing system.</p><p><b>METHODS</b>Concentration gradients of betaine were introduced into the sequencing system by taking the 5' terminal of Nogo-B cDNA (Am-Nogo-B) (G-C%=72%, without trinucleotide repeats) and 5' terminal of Huntingtin cDNA (Am-HTT) (G-C%=74%, with abundant CAG and CCG repeats) the results of sequencing were compared.</p><p><b>RESULTS</b>The optimum concentration of betaine for sequencing Am-Nogo-B has differed from that for Am-HTT. Result of sequencing Am-Nogo-B has achieved the best quality when the concentration of betaine was at 0.8-1.2 mol/L, whereas the result of sequencing Am-HTT obtained the best quality when the concentration of betaine was at 1.6 -2.4 mol/L. The results were reproducible.</p><p><b>CONCLUSION</b>G-C rich DNA with similar G-C% required different concentrations of betaine in the sequencing system due to base pair compositions. The sequencing system developed for improving the resolution of sequencing of G-C rich DNA with abundant trinucleotide repeats can be used as a reference for similar studies.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Betaine / Molecular Sequence Data / Base Sequence / Sequence Analysis, DNA / Trinucleotide Repeats / Huntingtin Protein / Genetics / Methods / Nerve Tissue Proteins Language: Chinese Journal: Chinese Journal of Medical Genetics Year: 2014 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Betaine / Molecular Sequence Data / Base Sequence / Sequence Analysis, DNA / Trinucleotide Repeats / Huntingtin Protein / Genetics / Methods / Nerve Tissue Proteins Language: Chinese Journal: Chinese Journal of Medical Genetics Year: 2014 Type: Article