For the initial primer validation gel, the gDNA was degraded, preventing any bands from forming in the gDNA lanes. The PCR mixture prepared for the gene-specific cDNA sample evaporated during amplification as well! Because of this, we couldn’t get any data on our designed primer. 🙁 However, we ended up being able to test it in a later experiment along with the validated primer and the primer didn’t anneal, so it’s a bit of a moot point anyways. Thanks for the question!
BLAST (Basic Local Alignment Search Tool) is a very large database in which information nucleotide & protein sequences can be searched and analyzed. The NCBI Conserved Domain database is a similar tool for analyzing conserved sequences in specific genes. In our case, we used these tools to find and analyze existing information on Rrm2. Thanks for the question!
Can you further explain the primer validation gel not working due to experimental error?
For the initial primer validation gel, the gDNA was degraded, preventing any bands from forming in the gDNA lanes. The PCR mixture prepared for the gene-specific cDNA sample evaporated during amplification as well! Because of this, we couldn’t get any data on our designed primer. 🙁 However, we ended up being able to test it in a later experiment along with the validated primer and the primer didn’t anneal, so it’s a bit of a moot point anyways. Thanks for the question!
What are BLAST and NBCI?
BLAST (Basic Local Alignment Search Tool) is a very large database in which information nucleotide & protein sequences can be searched and analyzed. The NCBI Conserved Domain database is a similar tool for analyzing conserved sequences in specific genes. In our case, we used these tools to find and analyze existing information on Rrm2. Thanks for the question!
What are the cDNA and gDNA?
The cDNA lane is coding DNA (without the introns included) and the gDNA is genomic DNA (with introns included). Thanks for the comment!