How do you choose the detergent concentration for protein purification?

hey i am using 2%OG and 1%DDM what concentration do i need to use with DM? is there a formula to calculate? Why usualy using 1%DDM and 2% OG (what % is higher then the CMC?) thanks

31 December 2018 843 9 View

Where can i get information about PITY and PYES2 plasmids?

hey all i have inserted a gene into PITY and PYES2 in yeast(why both i will explain) the PITY is an integral plasmid (goes in the genome of the yeast)and the PYES2 isn't. i am looking to built...

10 November 2017 4,253 0 View

What are the plats for eby100- yeast cells with PYES2 plasmid?

hey i have EBY100- Saccharomyces cerevisiae yeast cells. with the plasmid PYES2 (the map is added) How do i make selective plats for it? what is the protocol? and what possitive and what negative...

10 November 2017 8,754 0 View

How to built Yeast library with mutations only in 2 helixes?

06 July 2017 7,666 3 View

How can i produce genomic analysis (DNA extract) with very low concentration of DNA from Yeast?

hey all i need to extract DNA from yeast cells. but how can i do it from one yeast or very low number of yeast cells. is there a special protocol for it? Thanks May

05 June 2017 4,234 10 View

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View