ExactStart™ Eukaryotic mRNA 5'- & 3'-RACE Kit
ExactStart™ Eukaryotic mRNA 5'- & 3'-RACE Kit
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Last modified: 2008-07-15 14:57:15
The ExactSTART™ Eukaryotic mRNA 5'- & 3'-RACE Kit* facilitates the precise identification of the 5' and 3’ ends of eukaryotic mRNAs that contain both a 5'-capped and a 3'-polyadenylated end (e.g., 5' m7GpppN—[A]n 3'). The kit uses sequential treatment with APex™ Heat-Labile Alkaline Phosphatase and Tobacco Acid Pyrophosphatase (TAP) to selectively 5’-tag only RNA with a 5'-cap structure. Subsequently, a T4 RNA Ligase-mediated (RLM) tagging process adds an RNA oligonuclotide, containing a PCR priming site, to the 5' end of these RNAs. The 3'-end of the RNAs is tagged by reverse transcription using an oligo(dT) primer with a PCR priming site at its 5'-end. Thus, only RNAs that have both a 5'-capped and a 3'-polyadenylated end, such as eukaryotic mRNAs, are tagged with PCR priming sites. RNAs that lack either or both of these features are generally excluded from the population of amplified double-strand cDNA (ds cDNA) produced by the kit. The 5'- and 3'-tagged cDNA is converted to ds cDNA and amplified by PCR for 5’ and/or 3’-RACE. Due to the process of tagging both the 5’ and 3’-ends of the RNA, there is a very strong selection for producing only full-length amplified ds cDNA.
Last modified: 2008-10-29 09:35:16
Last modified: 2008-07-15 14:57:15
