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Questions related to Next Generation Sequencing
I have carried out an exploratory factor analysis (PCA). Using factor scores resulting of such EFA, I have carried out a new factor analysis. The problems is that in the factor analysis of the...
15 September 2019 9,012 8 View
Dear All, could some please suggest proper PhiX concentration for multiplex Illumina sequencing with use of MiSeq kit v2 and Nextera XT. samples #1-11 are highly similar - around 50-56%GC sample...
08 September 2019 3,077 1 View
I have access to the patient's blood who has been diagnosed with the MEGDEL syndrome. and we need to perform the mRNA analysis (splicing) based on the mutation position. I would like to know...
02 September 2019 8,173 6 View
I would like to do next generation sequencing on cells sorted based on an intracellular stain. Is there a best practice for how to fix the cells such that my sequencing run is not compromise?
29 August 2019 7,972 2 View
I performed immunodetection of modified nucleotides (5mC and 8dG) on neuronal and astroglial primary culture, and I expected to observe the fluorescence signal only from nuclei, whereas the signal...
27 August 2019 7,061 1 View
Is q-PCR sensitive to Pseudogenes?
16 August 2019 3,038 2 View
Since I m new to NGS, I have this basic doubt. Kindly pour your knowledge on this regard.
06 August 2019 4,109 3 View
Hi, I would like to try to express a fusion protein of LAMP-1 and a fluorescent protein to direct the fluorescence to lysosomes. The fluorescent protein will be fused with the C-terminus of...
29 July 2019 5,976 1 View
Hi, I have an isolated species from environmental samples, what is the best way to identify it? I think using Sanger sequencing with total 16S will be enough, but I heard that NGS (shotgun method...
17 July 2019 7,454 6 View
I´ve been searching for controls in NGS but I´ve only find quality controls. So I´m in doubt if it isn't necessary the positive and negative control during the sequencing. Hope someone can help me
10 July 2019 7,577 1 View
Hi, I'm doing an analysis of the immuno repertoire en T cell from different patients and this in several conditions. How could I compare two sets of TCR sequences (each set are arround 100 000...
08 July 2019 1,476 7 View
CMAP is a wealth of data, but I'm finding the sheer volume of transcriptional signature data generated given a drug (e.g., /sig amitriptyline) very hard to work with. The CLUE website will use...
03 July 2019 372 0 View
82% coverage has already been obtained, with a good quality sequence data using PacBio. Do we necessarily need to fill up the gaps using short read sequencing on Illumina.
28 June 2019 8,074 3 View
My primary antibody is bet- galactosidase (biotin-conjugate) I am not getting primary Ab binding but all my slides are stained dark with secondary and non specific binding. Should I use mouse on...
25 June 2019 7,398 23 View
I am trying to amplify two genes, each ~2kb in length. I am looking for ways to do this at the stages of cDNA synthesis and PCR amplification. Primer design and thermocycle specifications do not...
13 June 2019 2,486 1 View
I have NGS data of TCR-a from probiotic and control group. I want to see if there are identical sequences between these 2 group. In order to do that, I want to find out what are these identical...
01 June 2019 8,024 6 View
I will perform a MethyLight assay with DNA of brain rats tissue. Wich control should I use to normalize de methylation percent of reaction? We found inventoried controls for humans, but not for rats.
17 May 2019 9,275 5 View
We have tried imaging the NMJs of adult drosophila legs before, whereby on anti-HRP was used as the only primary antibody. Several papers that I have read through have also mentioned anti-Dlg and...
09 May 2019 6,500 1 View
Hello everyone, I was dealing with library construction for low complexity region from human genome, And I also found that, the sequencing depth are obviously bias: which means, some reads were...
07 May 2019 3,725 11 View
I am doing a project on trying to stain and image the neuromuscular junctions of drosophila leg muscles in great detail (with the possibility of imaging the boutons as well). So I have got two...
06 May 2019 5,802 4 View
A research questionnaire is divided into four parts with 12, 15,12, 15 statements in each part. Is it justified to apply the factor analysis separately on each part?
30 April 2019 3,606 5 View
I am validating the screening of BRCA1 and BRCA2 genes by NGS (Ion Torrent). I would like to have more samples with pathogenic variants to compare with my samples and to confirm my validation...
22 April 2019 2,235 3 View
I'm planning on a gene expression analysis study of mosquito midgut in response to a virus. I want to know how long it will take for gene expression analysis using next generation sequencing. Or...
22 April 2019 8,579 3 View
I need to know that how can ve decide that this sampke needs 1 million reads so we have to load this much sample, where the otherone sampke needs 200k reads then it would be loaded the xyz.
21 April 2019 6,994 4 View