Has anyone faced background fluorescence problem of enzymes fluorogenic substrates?

While using 3 fluorogenic substrates derived from Coumarins in cellulose- and xylan-degrading enzymes screening, one of them (MUGlc) shows blue background fluorescence during enzyme assay. is...

31 December 2018 293 3 View

Need for Q1 Journal suggestion?

I am about to finish a manuscript about High-throughput screening of biocatalysts from metagenomic DNA library, I am in search of a Q1 journal with no limited words, pages or figures where I can...

03 April 2018 1,728 0 View

How do you define Novel Enzyme?

What are parameters that decide the novelty of an enzyme before characterize it? means If we have a protein sequence which shows certain similarity to a protein in the database, what percentage...

01 February 2018 5,553 11 View

Colony PCR and normal PCR?

I tried to do multiplex colony PCR, I pooled many clones into one tube and I run colony PCR and I did not get bands. To make sure that the problem is not my primers' design, I have extracted the...

31 December 2017 8,914 18 View

Solvent Accessible Surface Area SASA results interpretation?

How can we interpret SASA results when compare between Enzyme and Enzyme-ligand complex?

31 December 2017 2,416 0 View

How to decide the type of a protein from different families given in Blastx?

I have faced this with more than one gene. in the step of gene annotation of my NGS data I found in many case my enzyme of interest but I found that this enzyme is found in multispecies, it means...

11 December 2017 9,448 1 View

Would you please propose a software to compare NGS data to CDD?

After proceeding with Prodigal for Genes prediction we would able to blast it and find known genes in our data. Is it possible to find novel genes by comparing it to conserved domain database CDD?...

11 December 2017 3,882 7 View

Is there any substitute of FosmidMax DNA isolation Kit ?

Is there any manual way to isolate recombinant fosmid DNA from E.coli cells in the absence of isolation Kit?

03 April 2017 3,621 4 View

What is the maximum duration of storing bacterial library in 20% glycerol stock?

For how many time can we store bacteria in glycerol stock in -80 degree? Do we have to transfer the bacteria to fresh media with glycerol after an approximate duration? In addition to Freeze-Thaw...

11 December 2016 7,297 22 View

What is the best choice of Polymerase for large DNA PCR?

11 December 2016 8,669 30 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

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03 March 2021 5,360 2 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 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

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

Any research papers for extraction of nutrient deprivation on macroalgae? Analysed by HPLC-UV and GC/MS. Possibly from macroalgae genus Rhizoclonium?

I am doing my thesis on the nutrient deprivation on the macroalgae Rhizoclonium sp. I will be analysing the levels over a period of roughly 5 days by HPLC-UV and GC/MS. I will be using seawater as...

28 February 2021 6,477 1 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

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View