Do I need to prepare my nuclear lysis buffer hypo or hypertonic?

15 April 2013 1 505 Report

I am preparing nuclear lysis buffer and I have seen protocols with different NaCl concentrations included. Can someone explain whether I need 150mM NaCl or higher and why?

Xianfeng Wang

High salt helps lyse membranes and forces DNA into solution, according to Abcam.

Badges
Science method

More Ina Petrova's questions See All

This question was deleted ?

[DELETED]

19 February 2023 7,602 3 View

What statistical analysis will be appropriate with a small sample?

Hello, I conducted a pilot study with limited number of participants n=6. originally planned 20, but due to covid got less participants. I was checking A1C before and after telephone-based...

20 March 2021 6,622 8 View

RNA sequencing and plasmid copy number?

Hi everyone, I am in the very initial stages of planning an RNA sequencing experiment. I want to find out the differential gene expression in my bacterial strain of interest when exposed to...

12 June 2020 6,053 3 View

What can you do if extrapolated progression-free survival (PFS) is higher than extrapolated OS in partitioned survival model?

Hi everyone, I'm trying to do a partitioned survival analysis (PartSA) to model a cohort of patients directly from survival data in the trial and evaluate cost-effectiveness of two different...

10 November 2019 6,739 11 View

How do i align two protein structures in maestro and manually change amino acid residues in binding site according to the results?

how do i align two protein structures one with better resolution (but open form with no ligand) and another with transition state analogue residues (but worse resolution) and connect them this way...

08 July 2019 6,845 1 View

Had anyone established the presence of Lixisols in Europe?

I am interested in distribution of Acrisols and Lixisols. Also their genesis, chemical and morphological properties.

26 February 2019 1,431 0 View

What method do I need to use if I would like to see whether different forms of the same protein (mutant, WT) bind more or less to another protein?

I have two proteins of interest - one is GFP-tagged and one is mCherry tagged. Each protein has several different forms (wild-type, mutant). My hypothesis is that the wild-type (not the mutant)...

19 February 2019 5,946 3 View

Can anyone recommend the best statistical test for Sholl analysis and how to perform it?

I am trying to compare the Sholl analysis data I have between 4 conditions and I am trying to find out the best way to analyse it.

17 January 2019 6,514 0 View

Can iodoacetamide reduce disulfide bonds?

Hi guys, This is a question for experts on protein cystine / thiol chemistry. Can iodoacetamide (under any condition) cleave disulfide bonds of a protein without addition of a reducing agent...

26 November 2018 2,368 3 View

Do fixed Orthodontics reduce the oral pH?

An older study suggests that: See: R. Chatterjee, I. Kleinberg,Arch Oral Biol, 24 (1979), pp. 97–100 Is there any other research afirming or infirming this observation?

08 February 2018 5,397 3 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

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 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

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 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 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