SMART技术（SMART technology）.docVIP

SMART技术（SMART technology） SMART technology, The eukaryotic cell mRNA in the process there is a metaphor for the process of shoes to wear a hat , so the 3 end of mRNA with a Poly A, which is based on cDNA library by reverse transcriptase enzyme. But cDNA 5 sequences are different, how to obtain the full-length cDNA library, cDNA, how was obtained by reverse transcription of the trace mRNA how to get the full-length cDNA using the known fragment sequence (RACE), was a disturbing problem. It is common in double strand cDNA synthesis in two ends of the connecting joint, then amplified by a linker sequence known, or by using terminal transferase in double stranded cDNA 3 end and with a series of G or C (or A/T), then filled with sticky terminal was amplified using two known sequences. The. But there are some problems of these methods in different degree, such as the joint connection efficiency is very limited, will lead to the loss of information, and these methods require multiple enzyme limited samples of different processing needs, after repeated purification, will lose a lot of useful information, especially low abundance information in small samples. The correctness of the results will affect a large extent. In addition, since mRNA is partially degraded, it is difficult to determine whether the resulting cDNA is a full-length cDNA or a fragment of cDNA. The emergence of SMART technology is a new milestone. This is called the Switching Mechanism At end of the RNA Transcript 5 (SMART), is actually a very simple principle: 3 end with Oligo in the synthesis reaction of cDNA (dG) in advance to the SMART primers as the reverse transcriptase is synthesized with mRNA cDNA, mRNA at the 5 end of eukaryotic mRNA encountered the unique hat structure, namely the methylated G will continuously in cDNA terminal synthesis plus several (dC), SMART Oligo (dG) primers were paired with several C synthesis of cDNA highlight the end after the formation of extended template cDNA, reverse