How much chromatin Input control is used for Input library preparation for Sequencing?
CfDNA from cancer patients ofter show a trio of peaks products of mono-, di- and tri-nucleosome fragments. We usually repair and make liberaries directly without enriching for smallest peak (we...
03 April 2017 8,133 0 View
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 726 4 View
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 5,514 1 View
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 8,940 2 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 View
Hi all. As a beginner in ChIP-seq experiments, I hope you understand that the following questions might be somewhat basic. I am planning to perform ChIP-seq or MeDIP-seq analysis to investigate...
28 July 2024 6,938 1 View
Gene sequencing related trouble shooting
25 July 2024 4,149 2 View
In running two-dimensional gel electrophoresis on bacterial protein, some spots that appear to match a protein sequence have a significantly more acidic isoelectric point than the calculated pI....
24 July 2024 8,076 3 View
Hello researchers, Sorry for my stupid question. I am learning the QIIME2 workflow for analyzing some 16s amplicon NGS fastq data. I found a very nice paper with data and code public available...
20 July 2024 5,405 2 View
Hello all, I extracted RNA from my samples and performed RIP-seq. After annotating the genomic regions using R, I obtained promoters, exons, introns, and UTRs. Given that my samples consist of...
18 July 2024 1,579 2 View
Hi everyone, I am working on a hybridization chain reaction (HCR) and trying to visualize it with gel electrophoresis. The visualization of the HCR works but for the single components of this...
07 July 2024 9,413 4 View
I have DNA samples (extracted by using Qiagen kiit), the 260/280 ratio is consistence 2.1.. for most of the samples. Can I proceed for the whole genome sequencing with these values? NGS.
07 July 2024 3,751 4 View