Why there's no growth of HLA-A2 transformed cells on expected antibiotic plate?

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I am trying to resolve immune cytokines. Which is the right percentage of SDS resolving gel?

I wish to resolve IL-6 (mol wt= 21-26KDa), TNF- alpha (17.3-17.4 KDa), GMF-beta (16.7 KDa), GAPDH (36kDa) and PDL1 (40-70 KDa). I think 12.5% gel is good to go. Is it advisable or should I go for...

25 April 2024 9,798 0 View

How do i calculate cut off values in ELISA?

I am running ELISA for different biomarkers against meningitis. I need to know how to calculate cutoff values for it in ELISA. There is no negative control or positive control for these kits,...

07 April 2024 3,079 1 View

Need to justify choice of two deep learning models CNN and sae compared to other models in data analysis ?

I have selected two deep learning models CNN and sae for data analysis of a 1 d digitized data set. I need to justify choice of these two dl models in comparison to other dl and standard ml...

18 January 2024 6,800 1 View

Need to justify choice of two deep learning models CNN and sae compared to other models in data analysis ?

I have used two dl models CNN and sae for data analysis of 1 d digitized data set. I need to justify use of these two models in comparison to other dl and other models. I am using ga to optimize...

18 January 2024 9,816 0 View

Which primers should I use for random amplification of bacteria from CSF sample?

I am working on 16s rRNA gene sequencing. So for that I should do PCR with random primers. Suggest me with a primer that I should use, also the which universal buffer the ti can use and if...

14 December 2023 6,288 0 View

(How) Is it feasible to use a questionnaire (of closed ended questions) to explore the lived experiences of participants in a phenomenological study?

Is it usual to use the closed ended questions in a Google sheet before carrying out semi-structured interviews for collecting data in a phenomenological study? If it is possible how can it be...

08 December 2023 2,290 7 View

Can anyone suggests a free software for finding particle size ?

For the measurement of particle size from XRD TEM SEM etc

12 November 2023 8,821 2 View

Is there any alternate way to avoid perfusion using a perfusion buffer for isolation of liver cells for primary culture?

Currently, I am working on mouse or rat models' primary culture of liver cells (particularly hepatocytes). In most of the previous research, I found the protocol stressing the perfusion of the...

10 October 2023 3,685 0 View

Can anybody provide me a reference about, TAHARA task which is based on the flow solver created by Prof. Yusuke Tahara of Osaka Prefecture University?

Required to understand how to solve potential flow around a ship hull.

29 September 2023 6,949 0 View

In a screen-printed flexible PET-based sensor, there is a failure to demonstrate sensitivity in Electrochemical Impedance Spectroscopy (EIS)?

If you know the reason, kindly text me

23 September 2023 7,978 7 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

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 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

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

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View

What is the best way to drop a plasmid?

Please address the best way to drop a plasmid. Background: I have a "bait" plasmid resistant to kanamycin and a "prey" plasmid resistant to carbenicillin. After many rounds of streaking on...

25 July 2024 2,532 3 View

Isolation of PLV6-Bmal-Luc plasmid?

I am currently at a stage in my research where I will be using the pLV6-Bmal-luc plasmid (Addgene #: 68833). However, I am encountering some issues with the isolation of this plasmid and am unable...

23 July 2024 5,296 0 View

Junjie Shao

So, the plasmid is amp+ but Cl- according to the plate result?

Rehasri Selva Rajan

Yes Junjie Shao . But it shouldn't be like that. The growth was expected in plate 1

Michael J. Benedik

So it sounds like your plasmid is conferring amp/carb resistance but you no longer have chloramphenicol resistance. Please note that ampicillin and carbenicillin are essentially interchangeable for lab use.