Construction of plasmids suitable for in vitro synthesis of full-length mRNAs having a 3'-poly(A)+tail.
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.
Matsuo
Taisuke
T
Institute for Genome Research, University of Tokushima, Kuramotocho-3, Tokushima, 770-8503, Japan.
Obana
Eriko
E
Yamamoto
Takenori
T
Kakuhata
Rei
R
Yamazaki
Naoshi
N
Kataoka
Masatoshi
M
Baba
Yoshinobu
Y
Hori
Tomoshige
T
Shinohara
Yasuo
Y
D001483:Base Sequence; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D005822:Genetic Vectors / Q000235:genetics*; D007564:Japan; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards; D010957:Plasmids / Q000235:genetics*; D015347:RNA Probes / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics*; D012015:Reference Standards
D015347:RNA Probes; D012333:RNA, Messenger