If i perform cell lysis in 8M Urea should i still have inclusion bodies?

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Can i change buffer pH without changing precast gel pH?

Can i run a precast TBE native PAGE gel (made with tris pH 8.3) with 1x TBE running buffer of lower pH such as 7.0? Would the lower pH of the running buffer lower the gel pH if i pre-run the gel...

11 November 2020 3,377 4 View

How Can I Run RNA+protein Sample in Gel?

I am trying to do an EMSA with unlabeled RNA an some proteins. RNA is about 700nt long in-vitro transcribed and the 2 protein are about 30 and 60 kDa with PIs 6-6.5. One of the proteins is the...

03 September 2020 6,164 6 View

Do i need to learn programming for end-user bioinformatics?

I'm a molecular biologist, and i have a few projects coming up in transcriptomes and small RNA analysis. Can i get by without knowing any programming using user-friendly software such an Geneious...

10 May 2020 7,983 1 View

What is the best time-series modelling method for studying the impact of geopolitical risk in MENA on the oil market?

Hi, I am currently doing a thesis for my masters. My research paper is about studying the impact of geopolitical risk in Middle East and North Africa on oil prices. I wanted to run regressions...

19 July 2019 3,657 7 View

Concerns for Illumina sequencing for low GC content organism?

I want to go for whole genome sequencing using next gen Illumina but i have some problems. Firstly, the organism is a mollicute with a small genome and is an obligate plant parasite with low...

17 March 2019 8,278 4 View

Bacterial growth in cut shoot samples?

If i am preserving cut tree shoot samples in distilled water in the fridge with no antibacterial compound, will bacteria grow just on the base of the cut, or will it spread up the shoot in the...

21 July 2018 3,352 5 View

Anti-peptide polyclonal antibody Specificity?

Hello, I am designing to produce antibodies against 15aa peptides, these antibodies will be used for detection of a pathogen in plant. The problem is when i blast these 15aa sequences there are...

12 February 2018 6,743 3 View

Cell-free membrane protein expression, liposomes necessary?

Hi, I'm planning to express a membrane protein in a cell-free system, the Purefrex 2.0 one to be exact. I've tried absolutely everything and failed to do so in E.coli. They've put in their...

22 January 2018 1,200 6 View

Having Trouble expressing membrane Protein. Toxicity?

Hey, So i have a 39kDa Membrane Protein, cloned into pRSET C and transformed into C43, sequenced to makes sure everything is there and made a glycerol stock of. From that glycerol stock i start a...

15 December 2017 7,114 3 View

Am i supposed to see membrane protein band in sds page of cell lysate?

If i Lyse cell overexpressing a membrane protein with sonication in 20mM NaH2PO4 and 500mM Nacl, firstly can i load it as it is in sds page or do i have to add SDS loading buffer? Either way ,...

16 November 2017 10,052 8 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

I want to remove Urea from my protein sample?

I want to remove Urea from my protein (6kDa) sample. I did use : 1. Step down the Urea gradient (dialysis) in order to remove urea and refold my protein, but my protein still contains urea in the...

06 August 2024 1,350 4 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 View