If I will keep two terminator in one plasmid, is there any problem for transcription. Actually, I want to insert T7 terminator to pMV261 plasmid, it has already rrnBT1 terminator.
I am using pET28a vector for protein expression. This vector is containing two type of origin of replication i.e pBR322 Ori and f1 Ori. The f1 is the bacteriophage origin of replication which...
04 May 2018 2,759 5 View
How can I get the Chip-seq data of M. tuberculosis?
03 April 2017 930 0 View
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
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
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
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
What is the most suitable experimental setting to understand the consequences of a particular gene: (1) targeted degradation of the specific transcription factor (TF) of the gene, followed by...
04 August 2024 6,265 1 View
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
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
I'm a PhD Student. From Northwest Agriculture and Forestry University China. My Problem is this I start my work on bhlh79 transcription factor gene. I do my Y2H 5 time but the colonies appear on...
30 July 2024 1,412 8 View
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
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