Thắc mắc về DOP-PCR
- Thread starter NMT
- Start date
tran van giang
Senior Member
Mình cũng chưa làm cái này, có nghe nói thôi,
Bạn xem thêm đây này,
http://www.ncbi.nlm.nih.gov/pubmed/1639399
Bạn xem thêm đây này,
http://www.ncbi.nlm.nih.gov/pubmed/1639399
thanhtam_2601
Senior Member
Không biết bạn đã lên thớt chưa nhưng hy vọng câu trả lời này có thể giúp bạn.
Đây là 3 trường hợp có thể xảy ra khi thiết kế primer mà mình đã viết trước bây giờ ngại dịch ra chắc bạn đọc cũng có thể hiểu được.
Cloning a gene encoding A protein of the virus
The first, we find sequence of gene encoding A protein of the virus in GenBank. Three possibilities can occur.
Case 1: The sequence of RNA encoding A protein of the virus is published before. Base on completed coding sequence, we design specific primer to amplification gene encoding A protein.
Case 2: The sequence of RNA encoding A protein of the virus is very long. In Genbank, one fragment has sequence. After reverse transcription of mRNAs into cDNAs, we use four primers including two primers design in the known fragment, 5' RACE and 3' RACE.
Sequencing to detect completely sequence of cDNA encoding A protein.
Case 3: The sequence of gene encoding A protein of the virus is unknown. However, we found the sequence of A protein from other microorganism which are homology with the virus.
Comparing the sequence of A protein from other microorganism, we found conserved region. Base on conserved region sequence, we design degenerate primers to amplification one fragment gene encoding A protein and sequencing the fragment. The next step will follow case2.
Trường hợp bạn hỏi thuộc vào trường hợp thứ 3.
Đây là 3 trường hợp có thể xảy ra khi thiết kế primer mà mình đã viết trước bây giờ ngại dịch ra chắc bạn đọc cũng có thể hiểu được.
Cloning a gene encoding A protein of the virus
The first, we find sequence of gene encoding A protein of the virus in GenBank. Three possibilities can occur.
Case 1: The sequence of RNA encoding A protein of the virus is published before. Base on completed coding sequence, we design specific primer to amplification gene encoding A protein.
Case 2: The sequence of RNA encoding A protein of the virus is very long. In Genbank, one fragment has sequence. After reverse transcription of mRNAs into cDNAs, we use four primers including two primers design in the known fragment, 5' RACE and 3' RACE.
Sequencing to detect completely sequence of cDNA encoding A protein.
Case 3: The sequence of gene encoding A protein of the virus is unknown. However, we found the sequence of A protein from other microorganism which are homology with the virus.
Comparing the sequence of A protein from other microorganism, we found conserved region. Base on conserved region sequence, we design degenerate primers to amplification one fragment gene encoding A protein and sequencing the fragment. The next step will follow case2.
Trường hợp bạn hỏi thuộc vào trường hợp thứ 3.
