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- Title
Construction of plasmids suitable for in vitro synthesis of full-length mRNAs having a 3′-poly(A)+tail.
- Authors
Matsuo, Taisuke; Obana, Eriko; Yamamoto, Takenori; Kakuhata, Rei; Yamazaki, Naoshi; Kataoka, Masatoshi; Baba, Yoshinobu; Hori, Tomoshige; Shinohara, Yasuo
- Abstract
In vitro synthesized mRNAs are often used as calibrants in gene expression analysis. In the case of an analytical procedure for gene expression containing a process of reverse transcription such as microarray analysis, full-length mRNAs having a 3′-poly(A)+tail are required as calibrants. However, the preparation of full-length mRNAs having a 3′-poly(A)+tail by using ordinary plasmid vectors is difficult. In this study, we established two plasmid vectors in which the nucleotide sequence corresponding to the 3′-poly(A)+tail were included. By simply inserting full-length cDNAs lacking their 3′-poly(A)+tail into established plasmid vectors, full-length mRNAs having a 3′-poly(A)+tail were successfully prepared.
- Subjects
PLASMIDS; GENE expression; GENETIC vectors; MESSENGER RNA; NUCLEOTIDE sequence; NUCLEIC acid analysis; GENETIC regulation; MOLECULAR genetics; RNA
- Publication
Biotechnology Letters, 2009, Vol 31, Issue 2, p203
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-008-9847-6