지식나눔

dna 염색에서 dye per base pair ratio 계산방법

안녕하세요. 

제가 lambda dna를 yoyo로 염색하려고 하는데

많은 논문들에서 "dye per base pair ratio 1:10 " 비율로 

염색했다고 나옵니다. 

그런데 실험할땐 염색약과 dna의 농도로 계산해야 되는데.

이걸 몰수로 계산해서 하는걸까요??

생물분야를 많이 접해보지 않아서 확신이 않아서 도움요청드립니다.
  • dna 염색
  • lambda dna
  • yoyo
지식의 출발은 질문, 모든 지식의 완성은 답변! 
각 분야 한인연구자와 현업 전문가분들의 답변을 기다립니다.
답변 1
  • 답변

    배소현님의 답변

    첨부파일

    Question : How can I calculate the DNA bp: dye ratio?

    I have YOYO-1 dye and I want to add dye at 5:1 ratio. And I am using a plasmid of length 5411 bp. Please tell me how to calculate this ratio to get a volume of dye to be added to DNA of 1 microgram.

    Answer :
    If you mean molar ratio, you have to calculate the molar amount of 1 ug of 5411 bp DNA. It can be easily approximated with 660 g/mol per base pair, i.e. 1 ug of 5411 bp corresponds to 0.000001/(5411x660)=0.28 pmol. However, in your experiment you need to keep constant ratio of the dye per base pair, so the length is not important here and you just calculate 0.000001/660=1.5 nmol (of base pairs). So you need to add 7.5 nmol of the dye.

    Question : How can I calculate the DNA bp: dye ratio?

    I have YOYO-1 dye and I want to add dye at 5:1 ratio. And I am using a plasmid of length 5411 bp. Please tell me how to calculate this ratio to get a volume of dye to be added to DNA of 1 microgram.

    Answer :
    If you mean molar ratio, you have to calculate the molar amount of 1 ug of 5411 bp DNA. It can be easily approximated with 660 g/mol per base pair, i.e. 1 ug of 5411 bp corresponds to 0.000001/(5411x660)=0.28 pmol. However, in your experiment you need to keep constant ratio of the dye per base pair, so the length is not important here and you just calculate 0.000001/660=1.5 nmol (of base pairs). So you need to add 7.5 nmol of the dye.

    저도 찾아본 자료지만 답변 감사드립니다!ㅎ

    그러셨군요.. 크게 도움이 되지는 못했네요. 덕분에 저도 공부했습니다.