What is the effect/purpose of the sucrose and EDTA added to agarose in the original Ouchterlony Procedure?

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What is going wrong with PCR? Works perfectly for One-Step RT but fails if I try to divide into separate reactions?

Hi all, Hoping for some troubleshooting/advice. Currently setting up a virus typing assay for routine use. I have the assay working with good sensitivity (regularly amplifies samples that are...

09 June 2024 4,468 3 View

What is an injectable buffer for mice around pH10?

Hello, In the lab we use baytril (enrosite) as an antibacterial post surgery. It comes ready to inject for horses but we need to dilute it for our mice to give them the correct dose. The thing...

30 April 2024 2,845 0 View

Is Gravity Chromatography an alternative for Size Exclusion Chrom for Mass Spectrometry?

currently conducting a study trying to figure out the secondary metabolites from bacteria. Question #1: does it make sense if our flow goes from: extraction > TLC > SEC > TLC >...

23 April 2024 3,603 1 View

Does Enzyme Concentration Affect Km?

Hello, I am using Trypsin to determine its Km and Vmax values against casein from bovine milk. I know higher enzyme concentrations increase the Vmax value, but does more trypsin also affect the...

04 April 2024 8,608 3 View

Michaelis-Menten Equation and Substrate Concentration?

Hello, Background: I am trying to find the Vmax and Km values for 0.25% trypsin and the substrate casein. During optimization, I have tried different time points (1-15 minutes) and different...

01 April 2024 1,553 1 View

What do I do if my Bradford Reagent yields underestimated protein concentration for measuring trypsin?

I use 0.25% trypsin (2.5g/L) as a positive control in my protease assays. I'm trying to measure the stability of different proteases overtime and at different temps. When I measure the...

30 March 2024 8,491 0 View

Completely wrong plasmid sequence in transformants?

I have transformed NEB 5-alpha cells with a plasmid that I have pre-sequenced and confirmed it has the correct sequence. All I want to do is make more of this plasmid. However, when I transform...

04 December 2023 8,171 3 View

How do i transport of primary mouse hepatocytes ?

Hello, i have to transport some primary mouse hepatocytes in some 6 wells plates at 37 degrees celsius ideally. It's going to be a 45 min drive approximately. Does anyone have any ideas or...

06 November 2023 4,039 1 View

Does teaching infants sign language help the language acquisition process?

Research Design Project

10 September 2023 9,297 0 View

Any recommendations for Freshwater Fish of Costa Rica/Central America Literature Resources?

Howdy folks, I will be teaching a study abroad course in Costa Rica this summer (Caribbean side). My main research project with students will focus on freshwater turtles, but we anticipate...

06 February 2023 8,325 4 View

What precautions should be taken while handling S. aureus enterotoxin Type B in the lab?

I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...

07 August 2024 6,034 3 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 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 draw the five chelate rings of [Co(EDTA)]- on this picture?

Is this circle one of the rings?

03 August 2024 5,988 0 View

PBMC infection with virus?

In my study, I intend to infect PBMCs with SARS-CoV-2. After that, I will analyze NK cells by flow cytometry to see if their phenotype changes or if they show degranulation. After the infection, I...

01 August 2024 4,403 4 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

EDTA buffer solution?

Hello everyone! How can i prepare EDTA buffer solution (pH 8.2) to determine GSH levels? Thank you in advance.

29 July 2024 4,374 1 View

Do some Staphylococcus aureus strains have in vitro antimycobacterial activity ?

Some Staphylococcus aureus strains Inhibit the growth of Mycobacteria in Mueller Hinton Agar medium containing 10% OADC. Do some Staphylococcus aureus strains have in vitro antimycobacterial activity?

29 July 2024 10,023 2 View

Addition of EDTA during the synthesis of copper nanoparticles to prevent it from being oxidized?

I've attempted to use this method to synthesize copper nanoparticles. Copper nanoparticles can be synthesised using a variety of precursor materials. CuSO4, distilled water, NaOH, and EDTA are...

28 July 2024 8,027 3 View