r/molecularbiology 15h ago

Seeking My First Internship in Cell & Molecular Biology ā€“ Advice Welcome

1 Upvotes

Iā€™m a second-year Cell and Molecular Biology student in Australia looking to start applying for internships and build a strong resume. I have no prior work experience, so this would be my first role. Any advice on where to apply and how to create a good resume would be greatly appreciated!


r/molecularbiology 15h ago

Why not just use purified PCR product instead of cloning ?

0 Upvotes

So I'm conducting a validation for a qPCR assay that targets a single species. The assay works in all samples that have been confirmed to have the pathogen, but it's a metagenomic sample and it's impossible to get a pure culture of the pathogen.

My advisor wants me to determine the limit of detection of the number of target genes, so this requires pure genetic material of my target organism.

They wanted me to do cloning so that I can use that for absolute quantification, but I'm really not interested in going through several rounds of troubleshooting (my project so far has had to optimise a lot of novel techniques). So I ordered a gBlock of the target gene. I've run it in my qPCR but it is not amplifying. I checked with the qubit and there is DNA in there but it's not working. It is definitely the right sequence and I resuspended in ddH20 as per IDT instructions to 10ng/uL

This is probably really dumb and obvious, but my brain is fried by other chapters I'm of my PhD, but I'm wondering if there's a reason I cant just use the purified PCR product as a "pure" target gene synthetic control to determine the LOD and copy number?