Biological Resource Center, NITE (NBRC)

Q&A about Human cDNA Clones

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Announcement of the termination of DNA resources distribution services.

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Information for cDNA clones

Q1;
Can I order all human cDNA clones constructed in the NEDO human cDNA project?
A1;
We currently distribute Full-length human cDNA clones and Splicing-variant human cDNA clones that were constructed at the Helix Research Institute (HRI) and the Institute of Medical Science of the University of Tokyo (IMS-UT). You can search clones available from NBRC with NBRC Human cDNA clone search system on our website.
We do not provide the human cDNA clones constructed at Kazusa DNA Research Institute (KDRI). For details about KDRI clones, please refer to: http://www.kazusa.or.jp/NEDO/.
Q2;
What types of vectors were used to construct the NEDO human cDNA clones?
A2;
Two types of vectors, pME18S-FL3 and pUC19-FL3, were used in the NEDO human cDNA project. We provide you with information concerning the clones and vectors when we ship the requested human cDNA clones to you. Although each human cDNA insert is cloned between two Dra III sites, these sites were destroyed at the time of human cDNA insertion. Therefore, the Dra III sites cannot be used to take out the human cDNA insert from the vector. The clones can easily be amplified in E. coli in the presence of 50 µg/ml of Ampicillin, since both of the vectors contain a pUC replicon.
Q3;
What is the difference between the two vectors?
A3;
Most human cDNAs are cloned into pME18S-FL3, because it can be amplified both in E. coli and in culture cells as it contains a replication origin for cultured cells, a SRα promoter, 16S intron, and poly A signal derived from SV40 for gene expression. On the other hand, pUC19-FL3 can be amplified only in E. coli.
Q4;
I would like to know the primer sequences for sequencing human cDNA clones.
A4;
The following are appropriate primer sequences for pME18-FL3.
ME-FW (5'-TAC GGA AGT GTT ACT TCT GC-3')
ME-RV (5'-TGT GGG AGG TTT TTT CTC TA-3')
Several primers for M13-derived vectors or pUC-derived vectors are available for sequencing inserts in the pUC19-FL3 vector such as those shown below.
M13-M4 (5'-GTT TTC CCA GTC ACG AC-3')
M13-RV (5'-CAG GAA ACA GCA TTG AC-3')
Q5;
In what form will the human cDNA clones be sent to me?
A5;
All human cDNA clones will be sent out as a dried DNA pellet in a plastic tube. Just resuspend the pellet prior to use with an appropriate volume of ddH2O or TE buffer, pH 8.0.
Q6;
How much human cDNA will be provided?
A6;
We will send approximately 1µg of cDNA enclosed in a 1.5mL plastic tube.
Q7;
Can I also order E. coli cells harboring a human cDNA clone?
A7;
Unfortunately, we do not currently provide E. coli cells harboring human cDNA clones.

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 Culture Collection Division,  Biological Resource Center,  National Institute of Technology and Evaluation
Phone number:+81-438-20-5763
Address:2-5-8 Kazusakamatari, Kisarazu-shi, Chiba 2920818, Japan MAP
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