What are the effects of longer exposure to siRNA (for KD)?

1) I knocked down a protein and collected cells 24 h and 72 h after transfection with Lipofectamine (to note, 5 h after transfection medium was replaced with medium with FBS). 2) KD efficiency...

08 December 2020 9,850 1 View

Why is hard to produce antibodies that are specific to a protein?

Some antibodies reveal nonspecific bands when a western blot is performed. Why is hard to find a good antibody for a protein? Antibodies produced using as the immunogen a synthetic peptide or a...

03 December 2020 208 3 View

Why different antibodies for the same protein stain different places?

I tried two antibodies and one show stain in the cytoplasm and the other one in the nucleus. Why? (Blue - dapi; red - mitochondria; green - protein of interest) And why is hard to find a good...

24 November 2020 6,612 7 View

Could bone tissue be saved in 70% EtOH after decalcification before processing?

I got the mouse tibia and femur tissue harvested, fixed with formalin 48h and decalcified with 14% EDTA 72h before paraffin embedding. This protocol works well for both HE and IHC. Now, for the...

18 October 2020 5,318 1 View

Why are G-quadruplexes a thing? Why not A, T, or C-quadruplex?

Guanine-rich DNA sequences can fold into four-stranded, noncanonical secondary structures called G-quadruplexes (G4s). G4s were initially considered a structural curiosity, but recent evidence...

27 September 2020 9,105 6 View

How does methylene blue fluoresce under dark UV?

Will it show as blue color under dark UV light (say 390 nm to 400 nm range) ? Does it degrade over time under long exposure of UV light (i.e. losing its fluorescein characters) ? What...

23 September 2020 9,752 2 View

My citrate buffer for IHC turned light yellow after boilded 30min with tissue slice. Is it normal?

My citrate buffer for IHC turned light yellow after boilded 30min with tissue slice. I have adjuct the PH to 6 and buffer was fresh made. I just wondering why it turned yellow after boiled with...

09 September 2020 5,357 1 View

Can I only use Matlab-Simulink to model the network consensus problem of a multi-agent system in DC micro-grids?

Hello everyone,recently I have met some problems in my study.I want to achieve a multi-agent system in DC micro-grids,but I never used java so that I can't use Jade to achieve it. Can I only use...

07 September 2020 6,342 2 View

How to avoid washing cells out during the changing medium step?

I'm Culturing adult mice cardiomyoctes and during changing medium step the cells seems washed away. Any tips about how to avoid loosing cells during washing step?

02 September 2020 4,788 4 View

How to gain the access to image recognition?

I am graduate from other major, and now working in computer science and technology, my roomates are mostly researching on image recognition. I want to switch to it and learn how to cooperate with...

21 August 2020 2,888 9 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

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

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

Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...

17 February 2021 1,130 6 View

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17 February 2021 4,217 3 View