Gene sequencing problem - Getting a non gene of interest (Dec/19/2006 )
I have the weirdest results from gene sequencing.
I did not get my own gene of interest. Instead of that, I got nearly 100% of similarity of another and totally different bacteria after blasting it.
How can it happen? I did confirmed my genes for twice using colony pcr. I did restriction digest. Everything seems to be fine. Until now.
How can a different gene being cloned even after confirmation? Got the same thing after repeating few times for gene sequencing.
Argh!!! What's going on? SOrry about that and thanks alot for reading and giving help.
You should probably include more information. What are you amplifying? For example, if it is 16S PCR, it would be easily to contaminate since its a universal gene. Nevertheless, unless the sequencing unit has screwed up with sending you the results, it appears as though you have amplified a gene that is not what you wanted. Sequence the PCR product directly to see what you get.
I did not get my own gene of interest. Instead of that, I got nearly 100% of similarity of another and totally different bacteria after blasting it.
How can it happen? I did confirmed my genes for twice using colony pcr. I did restriction digest. Everything seems to be fine. Until now.
How can a different gene being cloned even after confirmation? Got the same thing after repeating few times for gene sequencing.
Argh!!! What's going on? SOrry about that and thanks alot for reading and giving help.