Help with Mold Indentification

[pkfamily]

Hey all. I'm new to this forum and I was hoping someone could help me with my mold project. My goal is to be able to grow mold I obtain from a source (rotten tomatoes or bread) and then sequence its DNA followed by a genome comparison to see what organism it is.

 

My problem lies with the sequencing of the mold DNA. Since I obtained the isolated mold from nature, I don't know what primer sequence to create in order to sequence its DNA.

 

 

Thanks.

[CharonY]

For identification you do not sequence the genome, just diagnostic stretches. Typically rRNA genes are amplified and sequenced. Note that normally sequencing is not done on the chromosome either. Depending on system one would use sequencing vectors or add linkers to the fragments to be sequenced, for example. For these elements you would use standard sequencing primers.

[pkfamily]

ok. How would I amplify the rRNA genes? I assume after I amplify the rRNA gene, I can insert it in a plasmid like TOPO and then send that in for sequencing.

Edited July 31, 2013 by pkfamily

[CharonY]

PCR using appropriate primer for fungal species.

[pkfamily]

How do I create the primer if I don't know what the fungal species is?

[CharonY]

You use conserved regions that are not specific to a specific species. You can easily find them with a quick lit search.

[pkfamily]

Thank you. I will do that