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지식나눔

지식나눔

Primer extension analysis

2006-06-20 org.kosen.entty.User@168f05a8 최동규(cdkcbk)
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뭔지를 몰라서 그래요^^ 수업 내용 중에 있었는데 모르겠네요.. 해결 좀 해 주세요^^*
  • Primer
  • extension
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    2006-06-20
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    Primer extension analysis is utilized to quantitate mRNA levels, and to detect low abundance mRNA species. In addition, primer extension analysis can also be utilized to map the 5'-end of transcripts to determine the exact start site(s) for transcription. A specific end-labeled primer is hybridized to the complementary region of the RNA. This primer is then utilized by reverse transcriptase to synthesize a complementary DNA molecule (cDNA) using the RNA as the template. The resulting cDNA is subsequently analyzed on a denaturing polyacrylamide gel. The length of the cDNA reflects the number of bases between the labeled nucleotide of the primer and the 5'-end of the RNA; the quantity of cDNA product is proportional to the amount of input target RNA. The ideal primer is a short single-stranded DNA fragment that is complementary to a region within the 5'-end of the transcript to be analyzed and is approximately 20-40 nucleotides in length. Extension products shorter than 150 nucleotides allow optimal resolution. Primer extension is a very sensitive technique; as little as 2pg of transcript can be detected, which corresponds to approximately 1 RNA molecule per cell (1). The versatility of this method allows primer extension to be used to address fundamental quantitative and qualitative questions about individual genes. >뭔지를 몰라서 그래요^^ >수업 내용 중에 있었는데 모르겠네요.. >해결 좀 해 주세요^^*
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    Primer extension analysis is utilized to quantitate mRNA levels, and to detect low abundance mRNA species. In addition, primer extension analysis can also be utilized to map the 5'-end of transcripts to determine the exact start site(s) for transcription. A specific end-labeled primer is hybridized to the complementary region of the RNA. This primer is then utilized by reverse transcriptase to synthesize a complementary DNA molecule (cDNA) using the RNA as the template. The resulting cDNA is subsequently analyzed on a denaturing polyacrylamide gel. The length of the cDNA reflects the number of bases between the labeled nucleotide of the primer and the 5'-end of the RNA; the quantity of cDNA product is proportional to the amount of input target RNA. The ideal primer is a short single-stranded DNA fragment that is complementary to a region within the 5'-end of the transcript to be analyzed and is approximately 20-40 nucleotides in length. Extension products shorter than 150 nucleotides allow optimal resolution. Primer extension is a very sensitive technique; as little as 2pg of transcript can be detected, which corresponds to approximately 1 RNA molecule per cell (1). The versatility of this method allows primer extension to be used to address fundamental quantitative and qualitative questions about individual genes. >뭔지를 몰라서 그래요^^ >수업 내용 중에 있었는데 모르겠네요.. >해결 좀 해 주세요^^*
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