what is a role transcription termination(TT) fragment in the Phil_s1 plasmid?
Can anyone help me with a heterologous protein purification problem from Pichia pastoris supernatant? I am trying to purify the 59.6 kDa protein from the supernatant (protein has secretion...
11 December 2017 9,344 0 View
Is it necessary to rate 260 to 280 plasmid after digested to be 1.8 then do transformation Pichia pastoris by electroporation? I use Phenol: chloroform: isoamyl alcohol, but I cant get suitable...
09 October 2017 3,542 4 View
What is the simple method extraction DNA from Pichia pastoris?
09 October 2017 7,475 0 View
What is the best way to purify the digested product?Is there another method other than ethanol purification?
07 August 2017 2,386 1 View
how can remove the endonuclease E. coli in plasmid extraction?
07 August 2017 972 4 View
how much DNA need for digest? The construction plasmid is 4700 ng/µL, I want linear it with Fermentas restriction enzyme (Stu I). and what is the best method for extract vector after digest?
07 August 2017 2,349 6 View
How much is complete DNA genome Pichia pastoris?
06 July 2017 6,090 0 View
How can extract DNA genomic with high purity in fungal with the CTAB method?
06 July 2017 7,005 6 View
I want to do a virtual clone. What do you recommend free software for the virtual clone?
05 June 2017 7,195 2 View
I want to Codon optimization of Thermobifidia fusca cel6B gene and its expression in Pichia pastoris. How can to do it?
31 December 2015 6,861 2 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
We assume this to be true. We also assume that the vacuum bomb is the latest version of explosives with an explosive power of a few to ten kg of TNT equivalent. It has the unique characteristic of...
04 August 2024 4,534 1 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