Monthly Archives: September 2012

Do you have ORFs from other species?

Answer: Currently, we only offer pre-made Human and Mouse ORF clones. We are in the process of producing full length ORF clones from other model animals such as rat. These full length ORF clones will be released to market in the near future. If you have full length ORF of any genes of interest from these model animals, please send us the information of these genes such as GenBank Accession or Nucleotide sequences. We will place them on the list of high priority Full length ORFs of genes for cloning into the vectors we offer. They will be charged with the same price as our other catalog O......
Posted in FAQs

I have done expression experiment, but there is no protein. Why?

Answer: A correct DNA sequence with a promoter does not guarantee you to get protein expressed. Many factors contribute to the success or failure of protein expression 1) Did you use the right type of expression vector for your expression host system, for instance our pReceiver-M01 vector is for mammalian cell expression and pReceiver-B01 is for bacterial expression? 2) If the protein is a membranous protein, does it need target sequence for proper localization? 3) Will the over-expressed protein be toxic to the host? 4) Will the protein be folded into a right three-dimensional structure......
Posted in FAQs

Have you done expression test of each gene?

Answer: All expression clones in pReceiever-B01 and pReceiver-WG02/WG03/WG04/WG05/WG09/WG16 have been tested for expression in E. coli or cell free protein production system made by Roche. Other expression clones with different features and for different host cell systems have not been tested for protein expression.  
Posted in FAQs

After I have done full-length sequencing of your clone, I found it is an alternative splicing form of the gene I am interested in, can you reclone and replace it?

Answer: We will make the best effort to clone the original splicing form and provide you with the replacement clone. However, we cannot guarantee that we will get the clone of a specific splicing variant.
Posted in FAQs

There is one nucleotide change in the sequence, and it encodes an amino acid for proper protein function as sited by many literatures, how will you handle this case?

Answer: Even though we cannot provide replacement clones for this type of cases, we do offer mutagenesis services to change any nucleotide compositions as requested for a nominal fee.
Posted in FAQs

Since you use PCR method to generate full length ORFs, how will you handle PCR induced mutation?

Answer: Our system incorporated a new high fidelity PCR composition that improves fidelity in generating full length ORFs by 10 fold. It is accomplished by reducing the non-specific binding of primers with template DNA at lower reaction temperature and by reducing the cycles needed for amplifications. We guarantee that there are no frame-shifting changes and no pre-mature stop codon in our full length ORF inserts compared with reference sequences. We will replace the defect clones or refund the charges if that occurs.  
Posted in FAQs

How do you explain the differences between your actual sequencing data and GenBank sequences?

Answer: The differences can originate from two potential sources or combination of the two: 1) naturally occurring events including polymorphisms, alternative splicings, inconsistencies in multiple copies/versions of the same genes resulting from annotation updates, sequencing errors and/or ambiguities; 2) artificially introduced changes by PCR cloning process. Very often, potential users of our clones compare our sequencing data with the sequence of only one reference transcript, and attribute any discrepancies to PCR introduced mutations. Through analysis of multiple clones of close to......
Posted in FAQs

How do you generate full length ORF to produce your clones?

Answer: We use PCR together with several patented technologies to maximize the fidelity of PCR to generate full length ORF (more information in ORF White Paper). We used more than 70 cDNA libraries of human or mice tissues as templates to generate full length ORFs.  
Posted in FAQs

I would like to sequence the ORFs, what the primer sequences can I use?

Answer: For the ORFEXPRESS™ Shuttle clones, the forwarding sequencing primer (from upstream of 5' end of ORF) is: 5'-CCCAGTCACGACGTTGTAAAACG, and the reverse sequencing primer (from downstream of 3' end of ORF) is: 5'-ATGGTCATAGCTGTTTCCTG. For the OmicsLink™ clones: you can find the primer sequences in our website.
Posted in FAQs

How do you do quality control of your clones?

Answer: During the production of an ORFEXPRESS™ shuttle clone, each full length ORF clone delivered to you is fully sequenced. Before the delivery of each clone, we perform additional QC by PCR with gene specific primers to ensure that you are receiving the highest quality from GeneCopoeia.  
Posted in FAQs