선택한 단어 수는 입니다.
최소 단어 이상 선택하여야 합니다.
최소 단어 이상 선택하여야 합니다.
선택한 단어 수는 30입니다.
최대 10 단어까지만 선택 가능합니다.
최대 10 단어까지만 선택 가능합니다.
제안 방법
- Another notable finding in this study is that we could identify PCR-amplified fragments on DNAs extracted from the paraffin-embedded samples and we can use paraffin- blocks for molecular genetic study. Most laboratories prefer to use fresh muscle samples than paraffin blocks, because DNA in paraffin blocks is partially degraded into smaller fragments due to the extensive and possibly deleterious interaction between fixative and nucleic acids.
- In an attempt to study the molecular nature of Korean MELAS patients and also to study further feasibility of using formalin-fixed and paraffin-embedded tissue to study mitochondrial DNA mutations, especially point mtDNA mutations, PCR (polymerase chain reaction)/RFLP (restriction fragment length polymorphism) analysis and mtDNA sequencing were performed. We could identify the 3243 (A to G) point mutation in three paraffin-embedded tissues of MELAS patients.
- 2. Schematic representation of the PCR amplification strategy to study the 3243 mutation in MELAS. The A to G transition mutation at 3243 will create a new Hae Ⅲ restriction enzyme site which is diagnostic for the 3243 MELAS mutation.
- The diagnosis of MELAS patients was made by clinical presentations, neurological examinations combined with enzyme histochemistry and characteristic electron microscopic findings (data not shown).
- With six DNAs, PCR amplifications were carried out in a volume of 100 jil using 20 μM of each dNTP, 2 units Taq DNA polymerase, 10 X Taq buffer, 30 pM of each primer (a forward primer, 3116-3134 and a reverse primer, 3353- 3333) and 50 n응 of template DNA. After a single predenaturation step (10 min, at 95℃), PCR was done by 1 min.
이론/모형
- Total DNA was extracted from paraffin blocks with xylene by a modified method of Love et al. (1993). Small amounts of tissues were cut from paraffin blocks using a sterile scalpel blade and incubated with 400㎕ xylene and 500 pl mineral oil for 2 h at room temperature under gentle agitation.
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