Sample contain mixed template.In that case how to read chromatogram file (Sanger sequencing)?
I am wondering how can it be possible to sequence mixed templates with Sanger sequencing.
Abhijeet Singh actually i am using universal primer to determine hosts of insects.In that case insects feeds on greater than one host contain mixed template.
Here you are talking about the sample, but the sequence file would have just one of many sequences.
One way is to clone the pcr product and re sequence clones when there will only be one sequence per clone. If you had only 2 sequences and one was known then you can subtract the known sequence at every base position and what is left is the second sequence. If one sequence is clearly weaker than the other then there are programs online that read secondary sequence and that may be worth a try
Mixed DNA is possible even from a single host if the host is heterozygote for the target DNA. Now for all such mixed bases we should technically obtain the IUPAC codes such as Y for A and T at same place. You can further clone this product to obtain one clear read and deduce another by substraction method as suggested by Paul above.
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