• Title/Summary/Keyword: FoLT-PCR

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Quadruplex Genotype Analysis at HumTH01, HumTPOX, HumCSF1PO and Amelogenin Loci by FoLT-PCR (FoLT-PCR에 의한 유전자형 (HumTH01, HumTPOX, HumCSF1PO & Amelogenin) 분석)

  • Lee, Yang-Han;Lim, Si-Keun;Kang, Pil-Won;Choi, Dong-Ho;Yoon, Song-Ro;Han, Myun-Soo
    • Analytical Science and Technology
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    • v.12 no.3
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    • pp.260-264
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    • 1999
  • A simple and rapid procedure, called FoLT-PCR(Formamide Low Temperature-Polymerase Chain Reaction) was applied to amplifying DNA directly from various forensic biological evidences including human blood, saliva, hair root, or semen without any DNA preparative steps. We added washing step with non-ionic detergent, 1% Triton X-100, and used Taq DNA polymerase instead of Tth DNA polymerase to amplify 3 STR loci and gender allele simultaneouly. Optimal concentration of formamide and annealing temperature were determined empirically to 8%(v/v), and $48^{\circ}C$ respectively. We also compared this method with standard PCR.

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Relationship between genetic mutations and diabetes in non-insulin dependent diabetic mellitus (NIDDM) (인슈린비의존성 당뇨병(NIDDM)에서 유전적 변이와 체질의학적 관계)

  • Kim, Cheorl-Ho;Lee, Tae-Kyun;Chong, Ji-Choen;Park, Won-Hwan;Kim, Yong-Ju;Kim, June-Ki;Park, Sun-Dong;Nam, Kyung-Soo;Kim, Yong Sung
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.2
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    • pp.141-148
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    • 1999
  • A simple and rapid FoLT(formamide low temperature)-PCR, whereby human genomic DNA from blood can be amplified without DNA preparative stps, is described using human insulin genes. By applicatin of FoLT-PCR in human insulin genes, intragenic polymorphism in non-coding regions of the human insulin gene was shown after amplification and analysis by restriction enzyme digestion. All nucleotide sequences were the same as the reported, and four necleotides, at 4 different positions were polymorphic, and polymorphic alleles ${\alpha}4$, ${\alpha}5$, ${\alpha}6$, and ${\beta}2$ were identified. The new alleles were originated from homologous recombination between the ${\alpha}1$ and ${\beta}1$ alleles, and the alleles were founded in heterozygotes only. Although allele ${\alpha}1$ was dominant, the new alleles and ${\beta}1$ were recessive. From the results, it was suggested that the new method of FoLT-PCR was highly applicable in genetic variation analysis.

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Diagnosis of Bovine Theileriosis by Direct PCR and Electrophoresis from Whole Blood Without DNA Extraction (DNA 추출없이 전혈을 이용한 PCR-전기영동법에 의한소의 타일레리아병 진단)

  • Seong Ho Kang;Sangmin Jang;Joon-Seok Chae;Yongseong Kim
    • Journal of the Korean Chemical Society
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    • v.47 no.2
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    • pp.127-132
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    • 2003
  • We have developed a direct polymerase chain reaction (PCR) and electrophoresis method for the diagnosis of bovine theileriosis from whole blood DNA analysis without DNA extraction. The technique empolyed a FoLT (formamide, low temp.) technique and was utilized in the diagnosis of bovine theileriosis. Formamide solubilize the blood cells, and the lowered incubation temperatures reduced protein coagulation. 100-200 nL of whole blood and PCR reagents were introduced directly into a PCR tube. After the amplification, the PCR product (816-bp DNA) was introduced into the electrophoresis system. The results of this analysis were consist with those obtained using purified DNA.