실험 Q&A Mol. Biol.-RNA > Small RNA Analysis
Internal control이 exogenous로 따로 넣어줄 경우
레벨1 diamedi (과기인)
Internal control이 exogenous와 endogenous로 나뉨 (참고: https://www.qiagen.com/ca/spotlight-pages/newsletters-and-magazines/articles/correct-interpretation-negative-results)
eature | Exogenous homologous | Exogenous heterologous | Endogenous |
Universal use in multiple assays | ✓ | ||
Serves as control for purification procedure | ✓ | ✓ | ✓ |
Differentiates purification errors from amplification errors | ✓ | ✓ | |
Template quantities are defined and consistent | ✓ | ✓ | |
Non-competitive internal control design | ✓ | ✓ |
Exogenous IC가 두가지:위 링크에 따르면:
1> homologous IC, targeted pathogen sequence와 같은 primer binding sites 가짐->an artificial template로 작용
target과 IC에 같은 프라이머를 쓰지만, sequence가 달라, 다른 프루브를 사용-pathogen과 IC amplicons 구분 가능.
2> Heterologous ICs:designed with their own primers and probe가 pathogen과 아예 binding site가 다름.
-allow for a design and setup that prevents competition for PCR components
-universal controls
-> 이에 따르면 human housekeeping gene과 동일한 gene에 대한 exogenous internal control은 넣어줄 필요가 없고, 오히려 농도가 높으면 PCR 반응을 저해하는게 맞나요>
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