I have library of plasmids and I am looking for a highthroughput method to sequence whole library instead of sequencing one by one. Could someone suggest any method?
So do you have minipreps from all clones you want to sequence? If this is the case, you should follow the DNA-seq protocol. But you say that you already have a library, so how did you construct it? Maybe you should be more explicit.
Dear Jose,
I dont have miniprep for each clone sepreatly. This library supposed to contain 721 different plasmids. mixture of 81 plasmids (different in two parts) are digested and then 9 pcr products are cloned in them. Final plasmids are different in 3 parts. I need to know sequence of these 3 fragments.
While you might be able to use DNA-Seq to sequence your library as suggested by Jose. The fact are not separate libaries and seem to be all in one pool, combined with the fact they are plasmids , means you are likely to have a very tough challenge in assembling the data afterwards. As you will not be able to determine which sequence has come from a particular plasmid if they are all pooled together
To perform high throughput sequencing you will need to barcode each plasmid in order to identify each one later on.
For me is no matter, which plasmid has which sequence. I just want to know how many different plasmids I have( to be somehow sure about size of library).
- Badges
- Science method