Did anyone attempt to shear the cfDNA prior to library prep?

03 April 2017 8,015 0 View

Can I take the standard deviations of average DCt?

I'm doing some qRTPCR data analysis on tumour samples and using macthed "normal" tissue as a control. Each normal and tumour was run in triplicate (technical). Now we want to use all the normal as...

05 June 2014 626 4 View

Is e-cadherin expressed (protein level) in HEK293 cells?

I read several papers and I found contrasting information regarding the expression of E-cadherin in HEK293 cells. I haven't run a WB as yet, but any personal experience will help.

03 April 2014 5,425 1 View

Fluorescence fading after transfection?

I'm having troubles maintaining my fluorescence, either RFPtag or ZsGreen, expression after a few days under selection with G418. I'm working with MCF7, T47D and NIH3T3. They transfect ok...not as...

06 July 2013 8,840 2 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