Riboprobe for invitro transcription _ one band on the gel?

17 September 2024 0 9K Report

I did the riboprobe with Sp6 polymerase after BamHI digestion of plasmid DNA . After running the gel, I got only one band (18s?). is my RNA degraded ? I need to Insitubhybridisation. Is my DNA is transcribed or not?

any one help me how to udnerstand .

Thanks

Badges
Science topic

Similar topics
Social Science
Media
Journalism

More La So's questions See All

Which pH range is allowed for subcutaneous pump administration?

Hello, We are designing a protocol to determine the effect of certain compounds in the health of rats, but our problems have certain instability issues when subjected to certain pHs. We were...

19 June 2024 5,950 1 View

What are novel cryptographic protocols that enhance email security?

what are novel cryptographic protocols that enhance email security?

13 June 2024 7,090 2 View

¿Alguien ha utilizado la Base De Datos ChemNETBase?

Me gustaría saber qué usos tiene la base y para que ha sifdo utilizada https://www.chemnetbase.com/

04 June 2024 1,112 1 View

Is there any sequence that needs to be added when synthesizing and cloning shRNA?

To create a cell line overexpressing a specific microRNA, I'm planning to synthesize and clone the shRNA sequence into a vector. Would it be sufficient to add only the termination sequence (TTTTT)...

27 May 2024 4,206 0 View

Gender medicine myocardial infarction sex difference what are?

sex difference in patients with myocardial infarction

17 May 2024 1,321 0 View

Fluorescence spectroscopy: signal from water?

In order to proceed with the quantification of an Azide Alkyne Cycloaddition reaction efficiency by using a fluorogenic probe with Cy5-DBCO, I first need to construct a calibration curve. The...

16 May 2024 6,266 5 View

Can I calculate the high and low spin of the material in Quantum espresso?

I would like to simulate the high and slow spin configuration of a metal oxide. I am using QE for it, but in my research, I found that this kind of calculus is not possible. Or if it is possible,...

15 May 2024 6,497 3 View

Blockchains performance comparison studies?

It is difficult, or even impossible, to find real performance metrics (transactions cost, time to transaction finality, transactions per second, etc.) on the different blockchains. In their...

13 May 2024 10,182 3 View

Is HK-2 reliable in its expression of kidney injury biomarkers?

I am trying to establish a model of hypoxia-reoxygenation in HK-2 cells in order to determine their expression of kidney injury biomarkers (e.g. KIM-1, NGAL, etc.) via qPCR. However, I am finding...

09 May 2024 9,223 0 View

How can I resolve the 'Redundant boundary conditions' error in my multiple-step Abaqus/Explicit simulation for my foot model?

Hello all, I am currently developing a multiple-step Explicit Foot model with the final goal to create a dynamic simulation of the stance phase. To do so, based on previous literature, my idea is...

02 May 2024 9,008 0 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

Does anyone know what might be causing the smeared bands on my western blot?

I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...

10 August 2024 7,480 3 View

Strugglling with m6A dot blot any suugesstion ?

I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...

10 August 2024 8,539 5 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 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

Does Anyone have expertise in in vitro transcription and RNA pull down assay?

I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.

07 August 2024 6,622 1 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

RNA later for the preservation of RNA in fecal samples at room temperature for one day (37°C)?

I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...

06 August 2024 1,367 3 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View