Encyclopedia of Experiments: Biology
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- 현미경 슬라이드에 버퍼 용액한 방울로 시작하고 그대로 웜을 물방울로 옮기십시오. 웜이 제자리에 있으면 보풀이없는 조직을 사용하여 버퍼를 흡수하고 제거하십시오.
최종 헹구기 후에 에탄올이 증발되면, 핵산 염료, DAPI를 포함하는 용액을 샘플에 추가합니다. DAPI는 DNA의 사소한 홈에서 아데닌 및 티민 기지에 우선적으로 결합합니다.
다음으로 시료의 유통 기한을 연장하기 위해 페이드 방지 방부제를 추가합니다. 그런 다음 파란색 필터가있는 형광 현미경을 사용하여 DAPI 바디를 시각화하는데, 이는 세포와 사이클 상태에 따라 단일 염색체, 자매 크로마티드 쌍 또는 전체 핵을 나타낼 수 있습니다.이 실험에서 우리는 DAPI 염색 된 자매 크로마티드 쌍을 난동구에 세어 카엔오르합의 테트라플로이드 균주를 식별합니다.
- 테트라플로이드 균주는 12개의 염색체 쌍을 가지고 있으며, 이는 DAPI 염색된 시체를 비료 난우세포로 계산하여 검증할 수 있습니다.
그런 다음 10 마이크로리터를 웜에 90% 에탄올을 떨어뜨리고 에탄올이 완전히 증발하는 것을 지켜보십시오. 증발이 끝나자마자 에탄올을 추가하고 증발하는 것을 지켜보는 과정을 반복합니다.
슬라이드의 장기 저장을 위해, 시판되 게 사용 가능한 또는 수제 안티 페이드에 벌레를 마운트. 다음 커버 슬립을 추가하고 매니큐어로 가장자리를 밀봉. 매니큐어가 건조 한 후, 슬라이드를 득점 할 수있다.
첫째, 가장 성숙한 수정되지 않은 난모를 찾아, 즉, 정자에 즉시 인접하고, 아직 정자 또는 자궁중 하나에 입력하지 않았습니다.
야생형 난모세포는 평균 6개의 DAPI 바디를 가지고 있습니다. 12DAPI 바디의 존재는 이 긴장에 있는 동물이 부분또는 완전한 tetraploids이다는 것을 표시합니다.
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