How does decreasing the height of sucrose cushion (1M, no gradient) effect ultracentrifugation / recovery?

19 March 2018 0 9K Report

I am aim to pellet ribosome footprints (ribosome bound to ~30nt mRNA fragment) by layering the cell lysate on 0.8ml of 1M sucrose cushion (no gradient) through ultracentrifugation at 71,000rpm for 3h.

I am wondering if it is okay to use lesser volume of the sucrose cushion (0.6ml) so that I can layer more volume of cell lysate.

In other words, if the height of the sucrose cushion decreases, will this affect precipitation of the footprints?

More Sonali Wonders's questions See All

Can anybody explain the term "Peak- Rating" in data obtained from LC/MS?

Data Acquisition detected more than 65000 metabolites with area maximum, RT-value and peak-rating. What is the significance of Peak Rating?

07 July 2024 6,643 2 View

How can we calculate the absolute electronegativity of NaLa(MoO4)2 compound?

I want to calculate the CBM and VBM from the band gap of NaLa(MoO4)2. But is unable find out the absolute electronegativity of NaLa(MoO4)2. Please help me with proper references How can I...

23 November 2023 9,701 1 View

What is the difference between H2DCFDA and CM-H2DCDFA? Can we use CM-H2DCDFA as a ROS indicator in confocal imaging of PLANT tissues?

Confocal imaging of cucumber leaf mesophyll cells by ROS I am using a 25 uM concentration with 30 minutes of incubation in the dark.

05 November 2023 8,032 1 View

Can any scientist provide COV434 cells for research work?

I need COV434 cells for toxicity study in human ovarian granulosa cells.

04 August 2023 5,182 0 View

Any studies done to evaluate how the grip properties (at dry, Wet and Oily) of the NBR coating vary with the chemical property of the latex type?

I've gone through a large number of literature but couldn't find relates rather saying crosslink density would affect it. Would you mind helping me with your knowledge to resolve this or help me...

25 July 2023 4,036 0 View

How to optimize crystallization condition if one complex gave multiple polymorphs once from a specific solvent but laterits form only one polymorph?

What experimental conditions or methods can be employed to induce the formation of multiple polymorphs in a cocrystal that previously exhibited various polymorphs with a specific solvent under...

04 July 2023 7,770 1 View

Are there any articles on systematic steps for developing a model based on qualitative and quantitative findings of a study ?

Hello, I am engaged in a research related to leishmaniasis and expect to develop a model based on the qualitative and quantitative findings of my study. But the challenge is the lack of literature...

29 June 2023 7,037 4 View

How to choose mobile phase in HPLC to separete the components having absorbace in similar wavelength?

Both components show maximum wavelength at 280 nm. How to calculate the concentration of both the components in that solution

14 June 2023 8,811 16 View

Different variables / data to access the impact of digital payments on the MSME sector in India?

Variables/data to access the impact of digital payments on the MSME sector in India.

26 February 2023 4,644 0 View

What is a way to isolate RPE cells from mouse eyes without retinal contamination?

I'm hoping to isolate RPE cells from mice eyes and perform PCR, however, contamination from the retina will interrupt my results. I'm wondering how people might have tried this in the past. I know...

11 December 2022 3,395 2 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 3 View

How does grain and grain boundary affect the ceramic when studying its dielectric properties?

I am not able to get good literature and the physics behind how first these grains and grain boundaries arises out of no where when we make a pellet to study its dielectric properties and then how...

07 August 2024 5,177 3 View

E.coli contamination in human RNA seq data ?

Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...

06 August 2024 807 3 View

How to increase protein lysate concentration?

I am trying to prepare samples for IP. The pellet size is good and I am using 1% Tx-100 and using a syringe to further lyse the cells and maintain protein-protein interactions. I keep having a...

01 August 2024 7,199 1 View

What is the best practice for resuspending cell pellets during competent cell preparation (both electro and comp)?

I'm fairly new to making homemade comp cells and have done so with some mild success. What is generally an acceptable way of resuspending comp cells during wash steps? I've been told to swirl the...

31 July 2024 8,309 0 View

Screening of EPS producer LAB?

if you have MRS broth already prepared, what is the manner you will follow for preparing hyper-sweet medium for screening of EPS producer LAB? what the concentration of saccharose or glucose you...

26 July 2024 6,606 0 View

Pink bacterial colonies?

I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...

22 July 2024 5,953 6 View

How to determine tannin content in fodder pellets?

22 July 2024 8,163 3 View

How can I determine if a drug binds to a ribosome using a Cryo-EM map?

Hello, I am searching for the binding position of a drug in a ribosome, based on previous work indicating it binds there. The resolution of the ribosome-drug Cryo-EM map is around 2.5 Å. I've been...

19 July 2024 2,155 0 View

Why my plant protein cannot resolubilize after TCA/Acetone extraction?

I am extracting proteins from plant samples and am currently attempting to use the TCA/Acetone extraction method. I am having trouble resolubilizing my sample after drying the pellet. I collected...

15 July 2024 9,025 3 View