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

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

Can I use a microplate reader to detect fluorescence?

This is my first time to do experiment using fluorescent proteins. And I am wondering a cheaper way to detect and directly measure relative fluorescence units as a way of determining...

07 August 2015 5,129 7 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Would fecal samples stored at -20C for over a year impact metagenomic analysis?

Dear All, I have access to some rodent fecal samples that have been stored at -20C instead of -80C. Anyone have experience with the impact of stowage condition on comparative metagenomic analysis...

01 March 2021 2,512 1 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View

How can I retrieve FASTA sequence from gene ID or UID of a Gene ? I have GeneBank IDs ?

I treid to use Entrez Batch, but it redirects me to NCBI site. Is there any script of Linux or Python through which I can download Fasta sequence of list of gene ID ?

21 February 2021 7,783 12 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

What is the significance of (isolated) high titers of IgG anti GD1a antibodies in an older patient with polyneuropathy?

Remote history of Campylobacter infection with reactive arthritis, and Spitz nevus (UMP). Arthropod exposure in Zika/ChikV endemic area prior to emergence of length dependent, sensory PN with...

18 February 2021 8,719 1 View

Repository for capture-seq raw reads?

I am a research tech in a wildlife genetics group and my PI and I are trying to figure out how/where to put raw reads in a repository. We have capture-seq NGS from Rapid Genomics. The post-doc who...

17 February 2021 7,575 2 View