Generating an efficient lentivirus/AAV-producing HEK293/FT or lentiviral particle?

Hi experts, I am trying to produce lentivirus/AAV-producing HEK293/FT? I have a few questions regarding the protocol and results. I followed multiple sources, but the one I really followed is...

06 July 2018 4,080 4 View

What is the best age of mice to start stereotaxic injection and the best time to see highest expression?

Hi Experts, Can you give me an advice on these critical conditions for stereotaxic injection in mice? 1. What is the best age to inject brain regions - striatum and substantia nigra? Based on my...

04 May 2018 2,486 3 View

Can I measure the DNA concentration with nanodrop after Chromatin Immunoprecipitation (ChIP)?

Hi Experts, I am trying to do ChIP qPCR assay. So I followed the protocol from EMD Millipore/Sigma Aldrich kit for ChIP assay. According to the protocol, at the end of ChIP, just use 2 uL of...

03 April 2018 7,732 6 View

Why do we transfect cell during proliferation and not during differentiation?

Hi Experts, I am trying to transfect LUHMES cells and I've seen different protocols for transfection. Before or after differentiation, which one is better? And why? I've tried transfecting the...

03 April 2018 5,921 4 View

How can I identify and eliminate my cell culture contamination?

Hi experts, I need your help in identifying this kind of contamination. It has been a pain in my experiment to see this unforeseen event. This appeared first during my transfection experiments....

08 September 2017 4,021 8 View

In Western blotting, what is your experience with Wet vs Dry Transfer Systems?

Hi experts, I need your advice when using Dry Transfer System for transferring proteins from gel to nitrocellulose membrane. Have you ever used home-made gels, nitrocellulose membrane, and filter...

06 July 2017 8,936 15 View

What is the problem in my submission of metagenomic sequences to NCBI SRA?

Hi, I recently upload raw sequence reads to NCBI Sequence Read Archive (SRA). I followed the steps: (1) Make BioProject, (2) Make BioSamples (3) Submit to SRA (Beta) using Aspera Connect. However,...

06 July 2016 2,616 3 View

Converting GI number to other Database ID, and extracting batch protein sequences?

Hi Experts, I need your help. I need an updated online tool that can convert IDs from 1 database ID to another. I've been using bioDBnet website. However, I'm not sure if this is well maintained...

01 February 2016 7,741 2 View

What are the practical advantages and disadvantages of different protein purification methods?

10 November 2015 9,182 3 View

How do I determine if a peptide is cationic or anionic?

Hello Experts, I am still a beginner in this field, and I want to understand if a certain DNA or amino acid sequence is cationic or anionic? Is there a way (not experimentally), using...

10 November 2015 8,224 10 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

What is the exact mechanism of the antimicrobial effect of local anesthetics?

The antimicrobial effect of local anesthetics has been known for more than 50 years, but it still seems to have an unclear mechanism of action. which bacteria? which concentration? etc etc. ....

27 February 2021 4,450 2 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View