As above
I want to use two promoters to express two gRNA in a single plasmid and wish them to have same number(i.e. two promoter with nearly same strength). I am not sure whether I should insert a...
05 June 2019 7,016 1 View
I want to redo a RNA extraction from the E.coli cells and I store the liquid culture (in TB) for two days. I am not sure whether I can still use them or I need to regrow some cells.
01 February 2019 7,930 2 View
I was expecting only ~110bps amplicon in my sample, but I see a very broad band. Attached is the result. 2 and 4 only contain the template, no primers. Also, I think my intensity is kind of low.
10 November 2018 6,825 0 View
I want to compare transcription of a gene both inserted in the genome and a plasmid. Should I extract the RNA in the exponential phase or stationary phase?
10 November 2018 2,068 11 View
I want to count the amount of crRNA expressed in a plasmid with Ecoli. I am not sure whether I can use a RT-qPCR to do it. The length is between 50 - 100bps.
08 September 2018 1,169 6 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
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
I am currently at a stage in my research where I will be using the pLV6-Bmal-luc plasmid (Addgene #: 68833). However, I am encountering some issues with the isolation of this plasmid and am unable...
23 July 2024 5,296 0 View
