Sanger sequencing (=chain terminator sequencing)
Sanger sequencing에서 사용하는 ddNTP. 이것은 3‘-OH가 없기 때문에 dNTP자리에 삽입될 경우 그 이후의 DNA elongation이 일어나지 않게 된다.
Sanger method의 sequencing 원리
Sanger method - dNTP뿐 아니라 ddNTP를 사용해 PCR을 수행한 후 다양한 길이의 PCR products의 3‘ nucleotide를 확인
Single Nucleotide Polymorphism (SNP)
DNA sequence variation occurring when a single nucleotide –A, T, C, G- in the genome (or other shared sequence) differs between members of a biological species or paired chromosomes in an individual.
Variations in the DNA sequences of humans can affect how humans develop diseases and respond to drugs, pathogens, vaccines, chemicals, and other agents.
3. Dye-termination sequencing
Utilizes labelling of the chain terminator
Each of the four dideoxynucleotide chain
terminators is labelled with fluorescent dyes
Now being used for the vast majority of
sequencing projects
Challenge: differences in the incorporation of the dye-labelled chain terminators
into the DNA fragment
Blast
The Basic Local Alignment Search Tool
Ⅰ. Introduction
1. 배지만들기
1900대 초에 식물의 일부분을 떼어내어 배양하려는 조직배양 시, 세포에 영양분을 공급하기 위한 환경을 만들어주기 위해 배지를 만들기 시작하였다. George와 Sherrington(1984)에 의하면 1957년 Skoog와 Miller가 배지 내 auxin과 cytokinin의 상호작용이 조직이나 기관 배양시 기
Mechanism
Figure shows the mechanism for GM2 ganglioside degradation by beta-hexosaminidase A.
The GM2 activator protein binds to GM2 ganglioside which is attached to the lysosome's membrane phospholipids.
It then forms a complex with beta-hexosaminidase A by binding to the alpha-subunit.
Once this complex is formed, it can hydrolyze the beta-1,4-linked N-acetylhexosamine residue trans