What is the teal colored portion?

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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

How do I get good bands after PCR of gDNA from bacterial cells?

I have 6 bacterial isolates, I have checked their band on agarose gel electrophoresis, I am getting a genomic band but when I am performing the PCR and after running the gel, I am not getting good...

21 May 2024 8,614 4 View

How to retain the structure after optimization using Gaussian 16?

Dear Experts I have optimized an organic molecule using DFT/B3lYP/ 6-311G using Gaussian16. However, after the optimization, it was found that some bonds were missing from the molecule's...

17 May 2024 9,828 1 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

How to add parameters for "atotypin" ligand type?

I am running docking by using Autodock 4.2 software. However, I am not able to run autodock at the final stage and this type of command error appears in the terminal, Unknown ligand atom type...

10 March 2024 4,317 2 View

Does my Vero cells look contaminated?

Hi guys, We have been trying to get our cultures back after identifying contamination in our previous cultures. We have cleaned everything and started from uninfected cell stocks, but I’m not...

20 February 2024 6,717 0 View

Is skin yellowness an numerical or ordinal variable?

I have a response variable called skin yellowness, which I will measure via a scored color chart, whereby 1 is pale yellow and 15 is orange. I'm not sure if this counts as an ordinal variable,...

11 August 2024 4,793 1 View

Which type of compound does lamda max of 218 indicate in a uv-vis spectrum of a partially purified compound through column and TLC?

A crude extract of fungal culture using EtOH was subjected to column and TLC and partially purified compound was obtained. UV vis spectrum of the compound/s has max absorbance at 218nm. The...

11 August 2024 9,801 2 View

Can you connect an HPLC to a Mass Spec only at a certain time point?

Can anyone explain this method? Especially the last statement where it says only at 1.5 to 2.5mins was the MS/MS connected to the UPLC. How is that possible, is it a feature in this specific...

11 August 2024 8,141 3 View

RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

08 August 2024 9,882 2 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

Hi, please what is the best tool on Aspen plus that can help me get the best inlet temperature and pressure for cryogenic distillation?

I am trying to recover liquid CO2 from a mixture of 0.6 CO2, 0.3 N2, and 0.1 O2. My aim is to recover about 99% liquid CO2 at the bottom of the column and make sure the amount of gaseous CO2...

06 August 2024 4,611 0 View

Could dyes amplify the spectrum of light to a specific wavelength?

I am interested to know the behavior of dyes toward light. Specifically, Blue dyes re-emit the spectrum, especially from the green zone (known as principal in LED lamps, and blue dyes are known...

05 August 2024 3,290 1 View

What is the problem with these tissue culture plants?

All plants are green but some of these plants becomes yellow. I did not found any reason. Please help me to find out the real problem.

01 August 2024 589 4 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

What Are the Best Alternatives to the MTT Assay for Assessing Cell Toxicity of Colored Herbal Extracts?

I am currently investigating the cytotoxicity of a series of herbal extracts, and like many studies, I have been using the MTT assay to evaluate cell viability. However, I am encountering a...

31 July 2024 193 4 View