| Project/Area Number |
25860104
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Environmental and hygienic pharmacy
|
|---|
| Research Institution | Nihon University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2013-04-01 – 2015-03-31
|
| Project Status |
Completed (Fiscal Year 2014)
|
| Budget Amount *help |
¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
|
|---|
| Keywords | 遺伝子検査 / 食品衛生 / 食品衛生学 |
|---|
| Outline of Final Research Achievements |
In PCR analysis of processed foods, DNA degradation is essential problem. We focused on a DNA-degraded process and developed suitable extraction and detection methods for degraded DNA. We estimated the degraded process of λDNA as model DNA by high temperature and pressure conditions, and found the linearity between logarithms of PCR detection rates and exposed temperatures or times. Our DNA extraction method based on alkaline SDS lysis and DNA binding to silica membrane could conveniently extract a large amount of DNA from degraded samples, and had almost the same performance of PCR detection as general DNA extractions. In PCR detection, the probes contained locked nucleic acids could make the PCR conditions with small amplicon size and improve sensitivity in degraded DNAs. Furthermore, we developed detection method of a short DNA, about 20 mer, by reconstruction reaction using the template DNA with dUTP in model DNAs, although improving sensitivity and applying to foods are remained.
|
