DAPI staining protocol for protoplast?

Hanggara Aji Sakti Mahambara Padma Negara @Hanggara_Negara2

22 August 2022 1 6K Report

Hi, I want to stain my protoplast using DAPI to validate the nucleus position after I PEG-transformed it with GFP plasmid. Is DAPI staining method for protoplast is the same as staining plant cells/tissue, or there are some adjustment/modification? I really appreciate if anyone can share their experience or provide me some articles with detailed method in doing this. Thank you so much in advance :)

Ivan Kong

By theory, DAPI should stain A-T-rich regions in DNA. By that, your assumption may work as well like how DAPI could stain mammalian cells. Give it a try and provide your feedback to here, maybe? Thanks.

Badges
Science topic

More Hanggara Aji Sakti Mahambara Padma Negara's questions See All

Reusing PPM solution?

Hi, I want to use plant preservative mixture (PPM) solution to sterilize my explants (from ex vitro culture) and I have some questions: is it recommend/not to use the PPM sterilization solution...

10 February 2023 7,696 2 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

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 do soil microflora interact with plant roots and influence plant nutrition, health, and productivity?

06 August 2024 9,618 3 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

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 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

Why don't d-orbitals split themselves, why does it take a ligand? why don't protons from ligand repel nucleus split d-orbitals?

why don't d-orbitals split themselves because of themselves without the presence of ligands? Electrons are indistinguishable. Why wouldn't it be more correct that protons from a ligand split the...

03 August 2024 3,589 3 View

Why doesn't chromium 2+ ion use all its d-orbitals to receive lone pairs from six waters in [Cr(H20)6)]3+?

I'm guessing it's because the ligand experiences too much electron repulsion or proton repulsion from the chromium to insert them close to the 3d-orbitals which are close to the metal nucleus. Is...

03 August 2024 1,370 1 View

How can I get my Granzyme B flow cytometry stain to be consistent?

I have used PE and PE-Dazzle 594 fluorochromes and have managed to get NK cells to properly show GranzymeB expression after 4 hr PMA/ionomycin stimulaton, but for some reason my CD8 cells in the...

01 August 2024 7,677 2 View