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Questions related to Sequencing
I want to fuse a protein (830 aa) with RFP. I have some linker sequences, but I'm not sure which linker is suitable? short or long? which sequence? this protein is a heat shock one and expresses...
06 June 2016 3,338 4 View
I have transformed a vector (pET19b SUMO) from the lab as it was only in small quantities. However, once I've transformed and sent some sample off for sequencing, I only get ~1000bp out of a...
06 June 2016 6,996 5 View
Message ID#68 Error: Error occurred while trying to set up a Blast Object from CGI context: CFastaReader: Near line 1, there's a line that doesn't look like plausible data, but it's not marked as...
06 June 2016 8,889 6 View
We are into bacterial community analysis. As stated above we want to draw a phylogenetic tree based on 16S rRNA genes. As we have read in some publications it is possible first to draw a scaffold...
06 June 2016 5,475 1 View
I want to digest the rat gDNA by Spe1 to do Southern blot. However, according to my result, there still some gDNA didn't digest well. I am afraid it would affect the hybridization. Here is my...
06 June 2016 4,847 3 View
Anyone know a sequencing genomic center in USA received DNA samples from overseas?
06 June 2016 9,726 3 View
I have been trying to study DNA binding of a yeast transcription factor under different growth conditions using ChIP. Positive control shows enrichment under a specific growth condition and that...
06 June 2016 6,274 5 View
Hi all, I've recently sequenced a gene region, PCR'd from genomic DNA extracted from a cell line. From this, I've identified two regions of sequence variation, both within introns. I now want to...
06 June 2016 3,696 2 View
it is the first time that i use this kit and my questions are : SequalPrep Normalization Plate Kit is a good option for sequencing? and can i use only a fragment of the plate?
05 May 2016 1,590 2 View
Hello everyone, I am wondering what the best inducible expression system is for mammalian cells. Various companies offer their own inducible expression system for mammalian cells, but is one...
05 May 2016 1,812 1 View
I'm using DESeq2 to evaluate the effect of drug on gene expression from subjects with varying numbers of samples (2-5). Would this be biased? design attached
05 May 2016 6,920 5 View
I want to incorporate the GFP at the target sequence in the genomic DNA inorder to get a fuison protein, employing CRISPR-Cas9. My question is, should the GFP sequence be inserted within the...
05 May 2016 5,155 2 View
I am working on a project for archea and I want to propose to use Crispr Cas editing genome. I want to know how can I choose the hairpin (crRNA?) using Crispr Cas of Sulfulobus islandicus or...
05 May 2016 3,992 2 View
Hi all, I anticipate that I am completely new and for the first time I have to do cloning of genes whose sequence is unknown (The final objective is indeed to sequence the gene). Since I am not...
05 May 2016 8,282 1 View
please, I need to design probe as negative control in my experiments and i need to use Lac Z but i dont know its sequence can anybody help me in this. I will use it in mouse and human cell...
05 May 2016 4,231 1 View
Hello, everyone. Recently, I did chip-seq (H3K4me1、H3K4me3、H3K27ac、H3K27me3、Pol II、P300) from 1×107 insect cells. But, I got poor yield (about 4ng ) as standard Illumina library preparation needs...
05 May 2016 6,699 6 View
I want to start with some CRISPR experiments (with the nickase system) and I am wondering what sequences I should use for a non-target control? I am probably overthinking this, but at the moment I...
05 May 2016 5,324 2 View
Use of CRISPR/Cas9 on SORBIDRAFT_01g047430 and design of gRNAs.
05 May 2016 8,446 3 View
i selected a sequence for modelling currently it is not having experimental structures but model available in protein model portal.Can i model it and publish?
05 May 2016 7,828 2 View
We could observe similar stop codons in matching NCBI sequences. Is it justified
05 May 2016 859 3 View
I am working on setting up a CRISPR/Cas9 experiment. I have successfully cloned my sgRNAs, but due to the size of my deletion, will need to use a vector template instead of an ultramer. The...
05 May 2016 8,281 3 View
Someone here have been using the Duke sequencing facility ( (https://dugsim.net/estimate_cost). What about the reads/cluster quality? Thanks.
05 May 2016 7,063 1 View
Hi all, Are you familiar with Bioedit in the visualization of the sequences? I am trying to read sequences coming from Sanger sequencing method and I was said that the "genuine" sequence is...
05 May 2016 2,513 3 View
I want to sequence some unknown protein excised from 1D-SDS-gel. Have anybody experienced with protein sequencing? Are there any good commercial protein sequencing services?
05 May 2016 4,536 1 View