Hi,
I sequenced the genome of a thermophilic bacteria using Illumina. There was a gap of 37kb that was missed during sequencing but I was able to fill that using sanger sequecing. This region seemed to be highly conserved but has 33genes that are designated as hypothetical proteins.
My problem is I am not able to sequence a gene which is about 3.8kb completely. With internal pair of primers i was able to get 1.9kb sequence but am not able to walk in using primers from neighboring genes which is strange as I know the genes are together.
Can anyone suggest me what could be the possibilities?
This region is probably not stable in this organism it seems to be susceptible to degradation during extraction.
What could be the reasons?
I sequenced the genome of a thermophilic bacteria using Illumina. There was a gap of 37kb that was missed during sequencing but I was able to fill that using sanger sequecing. This region seemed to be highly conserved but has 33genes that are designated as hypothetical proteins.
My problem is I am not able to sequence a gene which is about 3.8kb completely. With internal pair of primers i was able to get 1.9kb sequence but am not able to walk in using primers from neighboring genes which is strange as I know the genes are together.
Can anyone suggest me what could be the possibilities?
This region is probably not stable in this organism it seems to be susceptible to degradation during extraction.
What could be the reasons?
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