Why housekeeping gene is showing thinner band than targeted knocked down gene with siRNA after PCR?

03 November 2017 1 8K Report

We transfected MDA MB 231 with SignalSilence® p42 MAPK (Erk2) siRNA, and collected RNA with RNA isolation kit. After cDNA preparation, we set annealing temperature and PCR was done, considering GAPDH as a housekeeping gene. We faced problem that after agarose gel run, it was showing that Erk2 band is thicker than GAPDH. We have done this PCR repeated times, but observation was same. If you could suggest us about the possible reasons regarding this problem, it will be helpful for us. Supporting picture is attached.

Ajay Saini

Hi, there seems to be no attachment. Kindly attach the photograph

Badges
Science topic

More Paramita Gayen's questions See All

Why the bandgap of CH3NH3PbCl3 films are different for spin-coating method and electrospinning method?

The observed bandgaps are 3.8 eV and 3.4 eV for spin-coated and electrospun films, respectively. What are the possible reasons?

08 January 2021 5,646 6 View

How to calculate SNR and throughput from path gain in IEEE802.11ac standard?

In MATLAB path gain is calculated using following command. Gain(i,j) = mean(abs(PathGain)); %Getting path gain at every location How to calculate SNR and throuput from this? I have Ptx and Prx...

27 December 2020 7,175 3 View

PL efficiency is higher i.e. better crystalinity for undoped semiconducto than doped one, but SEM shows less defect in doped one.How to explain it?

Undoped semiconductor shows better crystallinity in terms of higher photoluminescence (PL) intensity, in comparison to gallium doped semiconductor. But SEM images show that undoped semiconductor...

17 June 2020 5,554 8 View

What is the reason behind the appearance of sub-bandgap after 10% gallium doping in the pure semiconductor. But why the sub-gap disappeared for 20%?

Sub bandgap is absent in undoped semiconductor. Sub-gap appeared for 10% Ga-doping and again disappeared for highly doped sample such as 20%.

17 June 2020 1,505 4 View

Can I refer COD (Crystallography Open Database) ID instead of ICDD PDF no. for XRD analysis?

Instead of ICDD PDF number, COD ID can be referred or not?

14 May 2019 4,042 8 View

Is shifting of xrd peaks obvious during the doping? What extent of XRD peak shifting(e.g. <1 degree) is acceptable?

I have doped Sn in CH3NH3PbI3. There is shifting of peakes towards the lower value of 2Theta degree.

10 March 2019 5,771 9 View

What is the best method employed to extract silk fibroin protein from cocoon?

i am planning to extract silk fibroin proteins from cocoon. can anyone please suggest me the easiest and less expensive way for extracting silk fibroin protein?

22 November 2017 399 6 View

Is there any chemical procedure to get InI2 or GaI2? And the names of solvents in which they are soluble?

18 April 2017 4,781 3 View

Is Indium dichloride (InCl2) soluble in HCl or HI?

10 April 2017 4,617 2 View

Is there a good Mechanical protocol for Brain tissue dissociation for single cell suspension ?

I am trying to do ATAC-Seq from frozen brain tissue samples. Until now all protocols are from cell lines. I am looking for a good enough mechanical protocol, i.e., not using any enzymes to...

19 April 2016 9,742 4 View

Looking for Materials with high thermal expansion coefficient and high temperature resistance?

03 March 2021 8,272 1 View

Protein-aptamer gel electrophoresis help, please??

Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...

03 March 2021 4,275 4 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

How to regulate SSCP temperature in simple PAGE?

I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...

02 March 2021 9,157 1 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View