Who can hep me to analyze WES data for runs of homozygosity?

Hello, we have performed a WES of a single human sample and I am interested to analyse the runs of homozygosity in this sample. Especially, I am interested to unterstand if segmental uniparental...

10 November 2018 7,445 2 View

Has someone experience using Lipofectamine Stem Transfection Reagent for transfection of human induced pulripotent stem cells (iPSC)?

Has someone experience using Lipofectamine Stem Transfection Reagent for transfection of human induced pulripotent stem cells (iPSC)?

06 July 2018 2,129 0 View

Does T7 Endonuclease need overlapping nucleotides?

We want to know if Cas9 in combination with a sgRNA was working and have performed a PCR in combination with a T7 Assay. Unfortunately, we could not detect any cleavage product for several sgRNAs....

11 December 2017 2,477 1 View

Which bioinformatic prediction tool could be used to predict the influence of indel mutations?

Hi everyone, which other prediction tools (in addition to Provean) could be used to predict the Impact of indel mutations? Thanks for your help! Andreas

06 July 2017 5,616 3 View

Is it possible to compare one sample with a control set of samples?

04 May 2017 7,165 2 View

What are the best fluorescence proteins for structured illumination microscopy?

Hello everyone, I want to do SIM with three different proteins in living cells. Therefore, I need three different fluorescence proteins with high photostability and high brightness? Could anyone...

02 March 2017 3,522 6 View

What is the definition of stable cell line?

02 March 2017 5,366 0 View

Is it possible to use two fluorescence protein tags to increase the brigthness?

Hello everyone, I want to do SIM with living cells and therefore I need fluorescence proteins with high brightness and high photostability. Therefore, I want to ask if someone has fused his/her...

02 March 2017 9,698 3 View

Experience with Bmt1 (restriction enzyme)?

Has anyone experiences with the retriction enzyme BmtI? Are there important specific points to know about this restriction enzyme?

09 October 2014 2,679 1 View

Can you answer my general questions about BiFC?

I am planing BiFC assays and before I want to start I have some general questions: What is the strongest BiFC system? At the moment I could not say anything about the strength of my...

04 May 2014 3,947 3 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

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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 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