by admin | Sep 26, 2012 | FAQs
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.
by admin | Sep 26, 2012 | FAQs
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 20,000 human genes, we found that vast majority of discrepancies can be clearly attributed to category 1), when comparing our sequence data with all transcripts and sequences in corresponding gene locus including genomic sequence, ESTs, other cDNA/mRNA transcripts. On average, we found that there are fewer discrepancies between the sequences of our ORF clones and corresponding genomic sequence than the sequences of public domain gene transcripts and corresponding genomic sequences. We certainly cannot entirely eliminate the possibility of mutations introduced by our PCR cloning process, but we believe they are rare occurrences. We offer services at a small fee for changing base nucleotide compositions using a proprietary non-PCR based mutagenesis kit.
by admin | Sep 26, 2012 | FAQs
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.
by admin | Sep 26, 2012 | FAQs
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.
by admin | Sep 26, 2012 | FAQs
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.
