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

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

Can an isolated protein domain give a single band in SDS-PAGE equivalent to its dimeric mol. wt. ?

The protein I'm working with forms a homodimer and is a 2-domain protein. I have cloned and purified one of its domains. However, when I run the domain on SDS-PAGE, I get a single band equivalent...

19 August 2020 7,893 14 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View

Can part of recombinant protein not be expressed?

Hello. I am currently trying to express a recombinant gene with Avitag at the N-terminal. I'm aiming at a final product that looks like this: Avitag-(target protein) The protein I use is already...

14 February 2021 2,039 4 View

Transformation into the Stbl3 chemo competent cells?

03 February 2021 1,111 5 View

What is the best way and technique for serial dilution of bactria to get accurate number of colony ?

Hello I am infecting cells with bacteria in a well and then I am doing serial dilution of this well 1 in 10 and culture the two last dilution on MHA agar I have difficulty in getting accurate...

26 January 2021 9,256 5 View

Good day, I have not been able to amplify the plasmid Frt-hspB1. Somebody could give an advice me?

I acquired the plasmid in Addgene, reconstituted the filter paper where the plasmid comes from with 30 microliters of TE buffer pH 8, immediately afterwards perform the transformation of E.coli...

25 January 2021 2,526 3 View

Has someone used TTC agar overlay method and can help me?

I have tried to use TTC agar overlay method for a few times to identify respiratory-deficient yeast which were affected by EthBr. The problem is that cells which should have stayed white also have...

22 January 2021 585 10 View

I am having trouble with cloning my gene. How can I fix this?

16 January 2021 6,566 8 View

What is the best way to differentiate L. rhamnosus and L. casei in plate co-cultivation?

I'm having troubles in the differentiation and quantitation of each bacteria population during co-cultivation. The morphologic characteristics are very similar and it's really difficult to count...

13 January 2021 1,550 1 View