Can i use taqman gene expressn assay to detect a house keeping gene then use another taqman assay with different dye for the same gene rest of samp ?

More Basma Ali's questions See All

Is beta actin Rna stable?

in RTqPCR. if i have my target gene expression at ct 33 and beta actin at ct 17..i repeated and that's always the ratio..could my rna be degraded but i have good ct for beta actin how?

08 September 2019 7,057 4 View

Is there a difference between Rna expressed from cell culture and that extracted frim tissues?

if i can able to detect the transcript cd36 from cells but not from brain tissue .'.expected to have it'. ciuld it have more secondary conformational chnges than from cells?..just IDK

08 September 2019 5,748 5 View

Regarding Morris water maze test, if mice showed thigmotaxis in most of hidden platform training days whether reveciving LPS r control?

Should i exclude them from analysis? does LPS iP injection increases Thigmotaxis behavior?

02 March 2019 7,313 3 View

Nuclear protein PGC 1 alpha ? best way to extract from mouse brain ?

i have been struggeling to xtract it by ripa buffer ..then itake supernatent and heat in1 :1 ratio with 2x sample buffer at 95 degrees for 5 mins ..do yu have any suggestions ? it' s large ptn.so...

11 December 2018 2,068 2 View

Tips for large number of samples homogenization for rna extraction?

iam extracting RNA from brain tissues that are stored in -80 .should i get them all at same time from -80 which starting to homogenize some of them on ice or get few sames out of the -80 everynow...

10 November 2018 4,251 1 View

If i add protein extract to sample buffer is there difference if i heated till 60 for 10 mins than at 95 for 5 mins?

10 mins at 60 degrees vs 5 min at 95 degrees

10 November 2018 805 7 View

Is it better to reverse transcripe 1or 2ug of sample from brain tissue?

brain tissue reverse transcription..

10 November 2018 2,356 0 View

I cant get protein signals using RIPA buffer for extraction of one brain hemisphere ? (myhous keeping ptn is cytoplasmic ptn beta actin ? no signal?

i doubt i have a problem in my extraction procedures .. i homogenize my brain in RIPA buffer NACl 150 mM , 25mM tris , SDS 0.1%, Sdium deoxycholate 1%, Triton x 1%, edta 5mM ... i use it in 1ul...

10 November 2018 5,984 6 View

Can i multiplex 2 genes and singplex the rest in same run and compare results?

i mean i will do qpcr i will multiplex my housekeeping taqman gene (pl) vic dye with another taqman gene and will do the rest in singleplex format ? can i compare other genes expression to...

10 November 2018 2,960 3 View

Does alzheimer's disease affect novality seeking behavior in mice? or it's all about memory retrieval?

modified Y maze , would be glad if you posted advices /papers

09 October 2018 7,318 0 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

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

What precautions should be taken while handling S. aureus enterotoxin Type B in the lab?

I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...

07 August 2024 6,034 3 View

How to improve plaques in Double Layer Agar Assay?

Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...

06 August 2024 5,801 4 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 did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Could dyes amplify the spectrum of light to a specific wavelength?

I am interested to know the behavior of dyes toward light. Specifically, Blue dyes re-emit the spectrum, especially from the green zone (known as principal in LED lamps, and blue dyes are known...

05 August 2024 3,290 1 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View