Biological Resource Center (NBRC) / Q&A about Human cDNA Clones

Q&A about Human cDNA Clones

Ordering information

Q1; What types of documents are required for order of cDNA clones?
A1; The Ordering Form should be sent to us when you order cDNA clones.
Additionally if you order clone(s) which require MTAs, two original signed Material Transfer Agreements (MTAs) should be sent to us by postal mail.
Q2; Can I place my order for cDNA clones by fax?
A2; Yes, we can accept your order by fax.
However, if the clones require the MTA, it will be regarded provisional and your original MTAs are required for the formal processing of your order. Please send the MTAs to us when placing an order by fax. Do not forget to send to us two original signed MTAs. Your ordered clones will not be shipped until we receive the original documents. Note that photocopies are not acceptable.
Q3; Why do you require MTAs even though the Ordering Form contains an agreement for ordering the biological materials?
A3; Submission of MTAs are required to meet the distribution conditions under the contracts with the cDNA depositor.
Q4; Why are two original signed MTAs required?
A4; One of MTA is for the cDNA clone recipient and the other for the depositor.
Q5; Can I download the Ordering Form and the MTA from the NITE website?
A5; Yes. Please refer "Ordering Information" of our website.
Q6; Can I pay for the ordered cDNA clones by cash or check?
A6; Unfortunately, we cannot accept payment by cash or check. We currently ask you to pay either by bank transfer or credit card.
Q7; How do I select the method for payment?
A7; If you are paying by credit card, please select the check box "Credit card" in the Payment Method column of the ordering form. We will send you a proforma invoice and a credit card payment form by e-mail after we receive your order. Please fill out the credit card payment form and send it back to us by fax at +81-438-52-2329.
If you are paying by bank transfer, we will send only a proforma invoice to you. Please make your payment to our bank account as indicated in the proforma invoice.
Q8; What types of credit cards are accepted for payment?
A8; We currently accept only Visa and MasterCard.
Q9; Do you notify me that you have received my payment?
A9; No. We ship your ordered human cDNA clone(s) without prior notification of your payment. If, however, you do not receive it in spite of your payment, please contact us by e-mail at: dna@nite.go.jp
Q10; How many days will it take to receive human cDNA clone(s) after I place an order?
A10; We will make every effort to ship your ordered human cDNA clone(s) within 5 days after receiving payment information. However, as mentioned above, when you order by fax, we will not ship clones until we receive your two original signed MTAs if they are required.
Q11; Can I designate the shipping method?
A11; Sorry, but we will ship all ordered human cDNA clone(s) by special delivery within Japan and Express Mailing Service (EMS) to overseas countries. Unfortunately, Fedex is not available.
When we ship your ordered human cDNA clone(s), we will inform it with EMS item number to you by email. You can trace the clone(s) with the number at followin website: http://www.post.japanpost.jp/int/ems/delivery/ems_search_en.html.
Q12; I would like to know the general procedure for ordering human cDNA clones.
A12; Please refer "Ordering Information" of our website.

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.