Plasmid - vectors
pBI121 is commonly used in Arabidopsis labs - you can order it from the ABRC - http://www.arabidopsis.org/servlets/TairObject?type=vector&id=300033
Does anyone know of a UK/EU supplier of pBI121?
I'd also like to know where to get A. tumefaciens strain GV3101. Did you have any success finding it?
The primer annealing temperature , after the gradient PCR analysis, for the target gene differs from the optimal annealing temperature in housekeeping gene (positive control). What to do ? The...
06 July 2019 7,192 4 View
After several protocols for gDNA isolation i have noticed that in all samples teh DNA is totally degraded. I have used several papers, some of them...
02 March 2017 9,339 7 View
Dear all, How to calculate the RP - resolving power of primer base on the band number on Gel? In the example figure (taken from one good paper) one primer has 7.12 RP. Can anyone tell me how...
08 September 2016 3,458 0 View
For the past two months I have been trying to analyse my AFLP regions on a 6% UREA PAGE GEL. The sample are selectively amplified with 4 set of primers.I have tried to optimize the protocols for...
11 December 2015 9,106 14 View
Dear all, I have transformed my Ecoli with a clone of size (14.3kb). Plasmid alone is about 13.9kb and insert of size 432bp. In 1:3 ratio I got around 200 colonies, negative control plate (only...
08 September 2014 2,289 5 View
I am in the middle of my project- expression of U-OMP16. I am very pressured for time, and cannot wait for a fresh sample of infected animal with brucellosis in order to isolate the Brucella...
11 December 2013 7,443 0 View
Monoclonal antibody
08 September 2013 9,397 0 View
Respected, Does anyone know how to construct a phylogenetic tree base on (PIC) and heterozygosity (H) values calculated for each SSR primer using Darwin software ? The PIC and heterozygosity...
01 January 1970 4,609 13 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
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 have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?
29 July 2024 3,229 1 View
Hello everyone, I am currently looking for tools to recovery viral genomes from bacterial genomes, not metagenomes. However, I have only found tools that are designed for retrieving and studying...
28 July 2024 8,953 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
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