How to prepare highly efficient (10^8) TOP10 competent cells?

11 April 2020 0 10K Report

Hello,

I used CaCl2 Ionue method to prepare TOP10 competent cells two times, but obtained 10^6 efficiency in each case unlike DH5a that i prepared before which was very efficient. The OD of my culture was 0,3. I believe that bacterial culture was dissolved in buffer very gently on ice. How can I increase the efficiency of TOP10?

More Bilge Yaylak's questions See All

Can we joining HARDOX to HARDOX by Friction Stir Welding ?

We are try to HARDOX weld with FSW ? Is it possible ? I'm curious your opinion can you share me?

26 March 2024 9,438 3 View

Could anyone recommend an efficient electroporation protocol for primary preadipocyte cells or SGBS cell line?

Hi all, We seek an efficient electroporation protocol for patient-derived preadipocytes and SGBS cells to perform co-transfection for CRISPR/Cas9. They are pretty sensitive and hard to transfect....

12 February 2024 4,856 0 View

How to calculate SOMO value in Gaussian program?

How can I calculate the SOMO value to be able to perform SOMO-HOMO calculations? When I do the HOMO LUMO calculations of the structure I'm working on (spin doublet), I see two values due to the...

12 April 2023 6,654 2 View

Anyone has a cell binding, internalization and/or uptake protocol using radiopharmaceuticals?

Hello! I am trying to start a new experiment using 68Ga and 177Lu labeled peptides on LnCaP and PC-3 cells to find out the binding, internalization and uptake values. First of all, the terms are...

21 November 2022 8,405 0 View

Differentially expressed proteins?

Hi everyone, I have a proteomics data and want to compare male-female mice protein expression levels. I need to find differentially expressed proteins between two group. Student's T test just...

05 October 2022 9,681 2 View

Anyone perform RIP by using Magne® HaloTag® Beads?

I am planning to use an overexpression strategy to create a Halo-tagged fusion RBP protein, then aimed to perform RIP strategy by using Magne® HaloTag® Beads to reveal hypothesized RNA: Protein...

14 April 2022 695 0 View

Can I store cell extract, which crosslinked with formaldehyde, at -80 degrees before lysis ?

I tried to perform RNA immunoprecipitation assay to reveal RNA partners of the desired protein. Can I store cell extract not the lysate- after cross-linking (1% FA, 10 min RT)- at -80 degrees? Best.

14 April 2022 321 1 View

Have you ever experienced PBS poly(butylene succinate) electrospinning?

PBS poly(butylene succinate) electrospinning process conditions.

03 March 2022 1,720 8 View

What test(s) do you recommend for non-parametric binary data?

We are working on a study where we compare pre and post-workshop answers from a group of subjects. Subjects answered a question before the workshop and answered the same question after the...

17 January 2021 3,056 6 View

Can any one suggest a way to analysis of translational potential of candidate circular RNAs?

I have differentially expressed circular RNA candidates. Are there any data, database or bioinformatical pipeline to extract circular RNAs can potentially be translated into proteins? Thank you!

10 April 2020 8,628 1 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

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 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

How to use Density Functional Theory to calculate carrier mobilities of solid system?

Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...

04 August 2024 8,894 1 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