Optimized protocol for Pichia pastoris?

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Why is there cell debris are coming after 2nd or 3rd passage of the cells???

Our lab is facing a difficult problem for almost a year. After thawing the cells are okay, but after 2nd or 3rd passage there are debris present between the cells. We use mycoplasma removed cells...

17 January 2021 5,629 6 View

Can you help me for generating chicken swarm optimization matlab code?

Multi-objective optimization through chicken swarm optimization technique. Both maximization and minimization problem.

31 December 2020 2,299 4 View

How to remove straight line from mosaic Landsat image?

I have used mosaic to new raster tool to join two aerial image from Landsat 8 of different path. However, raster files had color variation and after mosaic I got a straight line running from...

22 December 2020 4,640 4 View

Any good methods for double-stained markers quantifications from tissue sample using computer programs?

How can we best quantify double-stained markers when comparing diseased and control? For example, if we want to stain one marker of smooth muscle cells and compare the expression in diseased and...

21 December 2020 7,389 1 View

Membrane protein Expression Purification: Broad Asymmetric peak on size exclusion ?

Hello, I am expressing an integral membrane protein (MW 51kDa) in Hi5 insect cells. I initially purified the protein by Nickel-His, concentrated it and subjected it to Size Exclusion...

10 December 2020 2,781 5 View

How should I store the milk sample in RNAlater?

I want to extract RNA from human breast milk. How should I store the milk sample in RNAlater? And after RNA extraction how can I store the RNA?

01 December 2020 7,183 4 View

Can U suggest standard protocols of quantifying total phenolics & total flavonoids using gallic or tannic acid for the former and quercetin for later?

The problem is that everytime getting the flavonoids content much higher than phenolics which may be, but in rare occasion. In phenolics, I am adding100 ul sample with 900 ul dH2O with...

29 September 2020 4,993 6 View

What is the ideal Elution buffer composition for a His tag protein purification to prevent protein aggregation after freeze thaw ?

Hi All, Recently, I purified 8 proteins and stored at -20 with the elution buffer containing 20 mM Tris, 250 mM NaCl and 200 mM Imidazole. I autoclaved the buffer, so I do not know whether the...

25 September 2020 9,814 6 View

What is the use of having multi-wavelength absorption measurements in Aethalometer?

Aethalometers are used to estimate the light-absorbing aerosols or Black Carbon (BC) or Soot particles. There are widely used aethalometers which are measuring light absorption in multiple...

06 September 2020 1,050 3 View

Has anyone done magnetic target localization using Genetic Algorithm?

I am trying to model magnetic target localization in MATLAB using those algorithms. So essentially what I want to output is the position vector of the target (x, y, z) and the magnetic moment...

28 August 2020 9,811 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

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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

High aspect ratio hexahedral mesh optimization solver?

I am interested in evaluating mesh optimization solvers that converge to high aspect ratio unstructured hexahedral grids. In other words, I am interested in a solver that optimizes warpage,...

26 February 2021 7,279 2 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to interpolate from a time-series database ?

Hi, I am having trouble with a problem, in the field of Optimal Control and the generation of optimal time-series. Let's consider a system, whose dynamics are represented by dx/dt =...

24 February 2021 9,105 2 View