How to construct a human gene into a ecoli plasmid for protein expression?

Saravanan Sureshkumar @Saravanan-Sureshkumar

18 May 2020 0 2K Report

Dear all,

I would like to clone a gene into a ecoli/ mammalian cell so if any one can support me and help me how to do from scratch. i have a project to clone some human genes, for eg: i would like to clone a human ADA gene to a plasmid construct how can i do it.

I kindly request the support from you all..

Badges
Science topic

Similar topics
Social Psychology
Attitude

More Saravanan Sureshkumar's questions See All

Design problem in a Buck converter for charging a Battery?

Iam designing a buck converter of input 200V with an output V of 48V with current more than 10amp for charging a battery. Can we convert 200v to 48 v, What can be the design problems that i can...

02 April 2024 6,847 2 View

How to correctly match the capacitance of ferroelectric layer and MOSFET capacitance in a ncfet?

We are doing simulations of fdsoi ncfet in synopsis tcad can someone correctly explain how to match the ferroelectric capacitance and mos capacitance

06 March 2024 7,511 0 View

In chemical oxygen method after adding ferroin the colour is red immediately and not blue. What is going wrong?

I am following APHA Method for determination. Final volume is 7.5ml. Should i dilute before adding ferroin

25 October 2023 3,585 0 View

Physcology of pricing in luxury watch brands?

Overall view and also a comparison view of Audemars Piguet & Patek philippe

13 July 2023 4,115 2 View

What solution should be used for dissolving CeO2 powders? & How many hours required to get homogenous (with/without heat) solution (gel) for coating?

Dear Researchers, I have tried to dissolve CeO2 powder in to acetone using sol-gel method. However, the powder got precipitated and not dissolving. So give your suggestion to get solution for the...

17 February 2023 2,691 1 View

Death after administration of Diminazene aceturate in recumbent cattle with babesiosis?

what may be the reason death of animal immediately or shortly after Diminazene aceturate administration? What may be the alternate approach or drug?

08 February 2023 8,471 2 View

Can you provide/suggest me internet users behaviour datasets (benchmark)?

Expecting the dataset contains user search query, time of search, visited links with time, etc.

04 February 2023 8,285 0 View

How to create a spherical wall?

Dear All, I need to put DNA inside the spherical wall (wall radius is 5nm) without any boundary conditions (PBC). Hence, DNA will be confined inside a spherical wall during simulation. Is there...

05 May 2022 5,348 1 View

Hi All, There is any standards for using seaweed polysaccharides in pet food?

Suggest the standards for using prebiotics in pet foods

15 March 2022 8,395 12 View

How the deposition time important in chemical vapor deposition for dielectric (SiOx/SixNx) thin films?

During CVD process, we got thick film and deposition time 120 minutes.

24 November 2021 7,916 3 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

How effective is the Citi Bloc standard basket in enhancing the accuracy and comparability of international construction cost assessments?

Citi BLOC Standard Basket Definitions: A standardized unit representing a fixed basket of construction materials, labor, and equipment costs priced in various cities. Purpose: To create a common...

04 August 2024 8,997 1 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 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

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