How to produce retroviruses pseudotyped with SARS-CoV-2 spike?

08 June 2020 0 9K Report

I am using pNL4-3 Luc R-E- and full length spike or deltaCT spike to generate Pseudotyped virus. Unfortunately, I am facing problem to get pseudotype virus. My positive control with pNL4-3 Luc R-E- and VSV-G is working well.

How much amount of plasmid to be used for co-transfection? May I get detail SOP who has generated retroviruses pseudotyped with SARS-CoV-2 spike?

Badges
Science topic

More Mahesh Agarwal's questions See All

How to calculate theoretical Ki from the docking results?

I have used Schrödinger (Glide) for docking

13 July 2024 3,003 1 View

Why T cells are not activating?

After PBMC isolation, we are not able to activate and expand T cells.

21 May 2024 6,124 2 View

How to create a thin film of tungsten oxide nanoparticles on glass plate?

I've tried to prepare a thin film of tungsten oxide nanoparticles on glass plate with the help of following methods- 1. Nanoparticles were dispersed in Poly-ethylene glycol (PEG) and the...

16 May 2024 9,007 1 View

How to find Euler angles for orientation of Single crystals with respect to Lab Frame?

Hello everyone, I have a monoclinic C2 space group single crystal sample glued to a tenon plate. I know the surface normal direction and the edge plan of the crystal. Also, I know the XY plane of...

27 April 2024 5,955 2 View

UDF code for 2dof forced osciallation of cylinder?

Hello I am trying to simulate 2dof cylinder for VIV analysis and I used an FDM method but the cd and cl values i got are so high and the damping is not coming into picture i tried and modified the...

17 April 2024 4,597 0 View

Why are PBMCs not proliferating in mixed lymphocyte reaction?

I have isolated PBMC from human blood and have set up an MLR for 5 days. I have then stained with CFSE and the cells have not proliferated. Any help would be appreciated

14 April 2024 2,514 1 View

How many wild type of rodents for study of zoonotic diseases?

Please suggest how many rodents required for any study wild type rodents

04 April 2024 4,483 0 View

How to assess proliferation in Mixed lymphocyte reaction with PBMC?

Hi, I am doing a mixed lymphocyte reaction with isolated PBMCs. To assess proliferation I have tried using CFSE, however, I can only see one peak after day 5 and not multiple. Has anyone faced a...

26 March 2024 4,934 1 View

Arruda-Boyce(8-chain model) UMAT availability?

Can someone provide Arruda-Boyce(8-chain model) UMAT code for Abaqus?

12 March 2024 2,854 0 View

Is there any subroutine in Abaqus that I can use to delete the whole part?

I can delete the elements, but can we also delete the part when the strain in an element of the part exceeds some value?

06 March 2024 1,245 3 View

How can I prepare virus for a TEM or SEM imaging?

I have virus (viral hemorrhagic septicemia virus) in suspension and the experiment will not involve cells. What level of TCID50 is preferred?

11 August 2024 3,115 1 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

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 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

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 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