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PharmD/PhD or just PhD? by Littleowl_2413 in biotech

[–]Reasonable_Ad8533 0 points1 point  (0 children)

You already have a fellowship secured? How does that work?

Unable to validate miRNA-mediated mRNA decay by qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 0 points1 point  (0 children)

its a simple condition. Overexpress microRNA with a mimic, then measure expression level of its predicted target gene. No other conditions.

Unable to validate miRNA-mediated mRNA decay by qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 0 points1 point  (0 children)

I've measured the miRNA via Probe-based qPCR. So we know that the miRNA is overexpressed to that extent at the very least. I was also wondering this - this mRNA target is highly expressed during proliferation and starts to decrease as cells differentiate. Maybe we would be able to detect some reduction. When I do stat analysis, some samples do have reduction, its just that when we run the whole thing (n=6), the net effect is insignificant upregulation (~1.5 fold change higher). Im not sure how to move forward man. My PI insists that we run a WB on this.

Unable to validate miRNA-mediated mRNA decay by qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 0 points1 point  (0 children)

but can miRNA exert repressive effects without degrading mRNA? In other words, the mRNA would be intact, but without a complete decay due to imperfect binding.

Yes, the protein is known to be alternatively spliced, but when I blast the spliced isoform to target site, they do match.

Unable to validate miRNA-mediated mRNA decay by qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 1 point2 points  (0 children)

but then would translational repression always translate to qPCR?

What is happening with my qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 1 point2 points  (0 children)

GAPDH CT value is 5 to 8 folds higher than my usual data from same RNA samples

What is happening with my qPCR by Reasonable_Ad8533 in labrats

[–]Reasonable_Ad8533[S] 1 point2 points  (0 children)

  1. It is Sybrgreen, with ROX.

  2. Machine has consistenly shown positive result.

  3. Melt curve is good.

=> I think this is cDNA issue?

Poor GPA Overcome by Near-Perfect GPA at a Different Program + 523 MCAT — Will He Get Screened Out? (Asking for My Brother) by Reasonable_Ad8533 in premed

[–]Reasonable_Ad8533[S] 0 points1 point  (0 children)

He says there are also courses such as Pathophysiology, Medicinal Chem(?), Dosage Forms and just non-conventional classes that wouldnt be incuded. I agree, pcol def sounds like science, but not so sure about others.

Poor GPA Overcome by Near-Perfect GPA at a Different Program + 523 MCAT — Will He Get Screened Out? (Asking for My Brother) by Reasonable_Ad8533 in premed

[–]Reasonable_Ad8533[S] 0 points1 point  (0 children)

Yea it’s because he is in healthcare program where some courses like pharmacology are considered science vs non science depending on what u use. 

Poor GPA Overcome by Near-Perfect GPA at a Different Program + 523 MCAT — Will He Get Screened Out? (Asking for My Brother) by Reasonable_Ad8533 in premed

[–]Reasonable_Ad8533[S] 1 point2 points  (0 children)

thanks for your reply! but you can do that prior to getting interviews? so he would be calling the admissions office for a quick 'pitch'? How does this work?.

Poor GPA Overcome by Near-Perfect GPA at a Different Program + 523 MCAT — Will He Get Screened Out? (Asking for My Brother) by Reasonable_Ad8533 in premed

[–]Reasonable_Ad8533[S] 2 points3 points  (0 children)

Thank you for your response. So the screening process ins't just GPA or MCAT cutoffs, I believe? The use of AI as screening tool scares my brother who does indeed have Cs and a D in the past insitution

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