When a fragment of gene was cloned into a vector for sequencing, two scenarios may lead to change of the sequence. 1) Errors in the PCR amplification, and wrong fragments can be cloned into vectors. ABI promises that the sequencing accuracy of its instrument can reach over 98.5% within a certain range. 2) Because of the limitation of the accuracy of the instrument, it is difficult to avoid a base sequence error in a longer sequence. After confirming the accuracy of cloning, bidirectional sequencing can minimize the sequencing errors. If you want to get your most accurate sequence, it is necessary to carry out bidirectional sequencing. With simple unidirectional sequencing, we can not guarantee the complete accuracy of the measured sequence, which is determined by the accuracy of the instrument.
Articles in this section
- What information do you need to prepare a quote?
- If you cannot finish the project (molecular biology project), do I still have to pay?
- I only have the vector map but not the sequence. Can you still process my request?
- Do you sequence verify the clones you made?
- Why I can't cut this plasmid (with XbaI siteTCTAGA) with XbaI while I can cut other constructs with Xba I fine?
- Why is my gene sequence different from the standard sequence?
- We have verified the insertions in the samples before submitting orders. Why the sequencing result indicates an empty vector?
- If client does not know the restriction enzyme sites flanking the genes, nor the vector map, but could provide the gene sequence, and primer sequences to be used for sequencing confirmation, can we take the project?
- With what vectors do we ship bacstab & induction solution to the client?
- Why did my PCR reaction stop working?