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Questions related to Sequencing
I have sent my sample for dna sequencing, now need to compare my mutated sequence with my data base sequence. so the question os which one is query and which one is subject sequence?
06 June 2016 6,353 3 View
I have a 16S rRNA sequence that has 100% of similarity to a bacterial species (Brevibacterium halotolerans ) that is included in a taxonomic group (Bacillus subtilis group). Does that result mean...
06 June 2016 3,191 8 View
If the Restriction enzyme site is in reverse direction at 5' to 3' end, can enzyme cut that sequence?(RE sequence is in Dark) 5'-GATACCTTAAGGTCG-3' 3'-CTATGGAATTCCAGC-5'
06 June 2016 2,710 5 View
I have amplified the whole mitochondrial genome in 9 fragments and use 62 set of overlapping internal primers for sequencing. I face problems while sequencing. Each fragment is further divided...
06 June 2016 4,870 1 View
I am trying to insert a fragment in pHisGb1, but after sending the miniprep for sequencing, find out the vector is empty. Now how can I improve my ligation?
06 June 2016 426 3 View
Hi there,I have a known fragment of my gene and I need of the 5' and 3' end to assemble them and get a full length of my interest gene. I thing use Inverse PCR, but I have a question. Is it...
06 June 2016 4,438 2 View
Hello contemporaries, any biomedical informatics databases that compares virus genome strains showing the sequence differences via a simple click? (Im interested in comparing adenovrius 40/41...
06 June 2016 7,682 3 View
I have a multiplex PCR with 6 Fwd species specific primers and one universal reverse primer. I usually run single PCRs and after this I clean them with exosap, then cycle sequence each with each...
06 June 2016 9,616 3 View
I've been trying to submit a sequence at Genbank using BankIt. It's an HRP2 exon2 partial CDS of Plasmodium falciparum. Everytime I paste my sequence it says that my sequence contains vector...
06 June 2016 8,134 3 View
Do the sequences in reference databases contain ambiguous N bases? if yes then why is it advised to remove ambiguous bases in quality filtering before mapping to reference??
06 June 2016 6,398 3 View
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 3,320 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 6,978 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 8,785 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 5,459 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 4,828 3 View
Anyone know a sequencing genomic center in USA received DNA samples from overseas?
06 June 2016 9,705 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 6,258 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 3,681 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 1,570 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 1,793 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 6,902 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 5,135 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 3,973 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 8,266 1 View