How does quantification graph of Beta-actin shows upregulation in QRT-PCR, when RNA was used instead of c-DNA?

More Satyam Singh's questions See All

Please explain how the plastic input value should be considered from the true stress-strain curve for the bilinear elastoplastic material model ?

I am working on Abaqus/Explicit(Quasistatic ) for the deformation of the auxetic structure model. Please explain how the plastic input value should be considered from the true stress-strain curve...

05 August 2024 454 3 View

How to increase citation in Research Gate?

How to enhanced h-index in Research Gate?

04 August 2024 3,368 4 View

How to do wavelet transform of EXAFS data?

I want to do wavelet transform of some compounds for their comparative study? Kindly help me in this regard and provide with proper procedure.

31 July 2024 4,170 0 View

Following click reaction in cell lysates, protein is immobile and remains at the top of the gel in SDS-PAGE?

I am using CuBr/THPTA for a click reaction in total cell lysates. I am facing issues with my protein sample in non-reducing SDS-PAGE where it's not migrating properly and most of it remains at the...

29 July 2024 950 4 View

How to use energy flexibility in inventory modeling?

29 July 2024 2,192 5 View

I want to get updates regarding international law, international trade law, international institutions and human rights etc?

Please send me updates regarding the above-mentioned areas in the question mark?

28 July 2024 5,197 3 View

What is the principle/mechanism behind aging of carbon (graphite) containing refractory mix for isostatically pressed refractories?

The resin bonded carbon containing refractories are aged before use. How time of aging is determined? And mechanism behind aging.

23 July 2024 3,205 0 View

How do we pick data for determination of Validation Acceptance Criteria?

Hello, colleagues! There is commenting open for new upcoming edition of USP 1033. Validation target acceptance criteria is now different from what it used to be and it doesn't include Cpm....

23 July 2024 7,292 3 View

What are the frameworks or methodologies to examine written academic ELF?

What should be the frameworks or methodologies to examine written academic ELF? I want to explore the linguistic features of written ELF in research articles.

23 July 2024 4,800 1 View

How to remove cell debris after tissue dissociation?

I'm facing issues with the tumor dissociation protocol for GBM. I use collagenase/dispase with DNase to dissociate my tumor sample. However, even after incubating it for 60mins, I do not get a...

22 July 2024 412 3 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 3 View

Strugglling with m6A dot blot any suugesstion ?

I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...

10 August 2024 8,539 5 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

08 August 2024 9,882 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Does Anyone have expertise in in vitro transcription and RNA pull down assay?

I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.

07 August 2024 6,622 1 View

E.coli contamination in human RNA seq data ?

Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...

06 August 2024 807 3 View

RNA later for the preservation of RNA in fecal samples at room temperature for one day (37°C)?

I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...

06 August 2024 1,367 3 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Narumi Uno

Hi,

I think the attached results are incorrect because PCR amplification by DNA polymerase using RNA as template is not possible when using DNA primer.

Sven Reister

beta Actin has ~100 pseudo genes in human DNA that do resemble the cDNA sequence (AKA no introns), so what you see is DNA amplification. To get this sorted, you may consider to have additional gDNA removal steps during RNA preparation or within the RT step (some kits do contain gDNA removal)

Sven