How do I introduce UCOE region before the hCMV promoter in the plasmid?

More G V Girish's questions See All

Developing hCG, FSH, LH biosimilar ?

Hi, We into developing hCG, FSH, LH biosimilar expressing CHO clones. Want to collaborate, or seek consultancy from the experts in the field. regards,

08 September 2018 4,165 0 View

Immortal heterohybridoma line secreting monoclonal anti-D antibodies ?

If any scientist ihas developed a Immortal heterohybridoma line secreting monoclonal anti-D antibodies, we are keen to collaborate to take the research to clinical level for conducting the...

08 September 2018 4,702 0 View

TGE is working but not SGE in DG44 cells ?

Our lab is working on developing the stable gene expressing clone in DG44 cells,. After transfection, transient gene expression is working, after growing the cells in the media with out GHT and...

07 August 2018 7,281 3 View

Looking for retroviral vector system for transducing DG44 cells ?

Hi We are Looking for retroviral vector system for transducing DG44 cells with GOI to produce expressing clone. Suggestion, collaboration are welcome.

02 March 2018 6,526 1 View

What is the difference in running a E.Coli Fermentor and Mammalian CHO cell bioreactor ?

what is the difference in running a E.Coli Fermentor and Mammalian CHO cell bioreactor ?

11 December 2017 5,291 0 View

Want to learn the 5'-ITR and 3'-ITR sequence of the Sleeping beauty transposon ?

Want to learn/know the 5'-ITR and 3'-ITR sequence of the Sleeping beauty transposon for designing the experiments working with Sleeping beauty transposase.

10 November 2017 3,743 3 View

Expression improvement in0 DG44 CHO cells and NANA glycosylation ?

Working on the DG44 CHO cells for recombinant protein expression, we have been getting the low expression of 0.5ug/ml recombinant protein and also Gal-a(1,3)Gal glycosylation of protein....

08 September 2016 8,082 1 View

Why restriction enzyme Sap1 does not cut the circular plasmid ?

Am working with cloning, and plasmid has an Sap1 restriction site at Forward and reverse (purchased the cut plasmid from an company, they call it Electracloning) and the insert gene has forward...

06 July 2016 8,268 3 View

Want to express hCG (alfa hCG + beta hCG) using human albumin signal peptide ?

Beta hCG expression is not seen, took of native signal peptide (20aa) of beta-hCG and replaced with human albumin signal peptide (18aa). Still B-hCG expression is not seen in the HEK293 cells. We...

03 April 2016 6,444 2 View

Two hundred USD Award for designing the Plasmid ?

We are working with pDUO2 plasmid and after transfecting it getting low expression of protein. I want to place A2UCOE sequence [HNRPA2B1 and CBX3] before the promoter for better expression, if any...

06 July 2015 3,584 2 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 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

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 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

Mingzhong Chen

Use Gibson assembly from NEB.

G V Girish

Thanks Dr.Chen.

I am not a molecular biologist, have no knowledge about editing the plasmid, my experimental aim is to put HNRPA2B1-CBX3 into the pDUO2 plasmid. Want to learn from you, will you please help with the experimental design of editing the pDUO2 plasmid.

regards,