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Questions related to Sequencing
Currently conducting cDNA amplification during a Tag-Seq (RNA-Seq) protocol. In the gel attached, all wells with 'D' should be cDNA with smears, all others should not because they are controls...
14 January 2017 3,707 1 View
I want to have sequences of a list of genes of several bacterial species. Many of these genes are not available on NCBI and other sources. But the whole genomes of the bacteria are quite a lot...
12 January 2017 6,000 6 View
Hi Everyone, i have a Sequence of 16s rRNA, when i tried to generate CAP- Contig using BioEdit, am getting Multiple Contigs for a particular strain, but when i tried with some other strain am...
11 January 2017 639 2 View
I have searched the literature and NCBI to no avail. I'd appreciate your help!
11 January 2017 9,076 2 View
I extracted DNA from blood and made bisulfite conversion. I did PCR (without cloning) and then sanger sequencing. I have an amplicon of 255 pb in a UTR gene. I also did PCR for the untreated DNA...
10 January 2017 5,404 4 View
Hi , I am new with the Illumina sequencing and I got back the sequencing data. I am quite new with the terminology. Can you explain to me what it means that a project will be sequenced using...
10 January 2017 7,638 12 View
I use vent exo- polymerase for amplification the template below but it seem not get high yield. Could you give me some advice? 5’-A (CTG)10 A AAG TTA GAA CCT ATG-3’
10 January 2017 2,286 5 View
Hi, Im learning about next gen. seq. So, with NGS you shear up dna into fragments and when the computer tries to align by matching overlapping regions isnt there a possibility of the fragments...
09 January 2017 258 4 View
Hi all. I need to evaluate Polyphenol oxidase (PPO) protein content in olive fruit extracts by Western blot. So I'm searching also a housekeeping protein in Olea europea for normalization. Some...
09 January 2017 5,360 3 View
How do I edit a gene manually after sequencing?
08 January 2017 1,057 3 View
I want to find out conserved domains of diacyglycerol acyltransferase in several algal species. But I am having difficulty to get the dgat sequences of C renhardtii. I will appreciate any help..
08 January 2017 5,407 2 View
I have approx 3000base ssDNA (3'-5', synthesized by asymmetric PCR) and complementary RNA (5'-3' ; synthesized by T7-transcription). I want to prepare DNA-RNA helix (hybrid) in solution. I have...
05 January 2017 3,495 1 View
We are trying to FACS sort shh-responsive cells from injured mouse facial nerve and run RNA-seq on them. I have limited bench experience. Any advice or detailed protocols for these procedures? I...
28 December 2016 1,677 2 View
I'm relatively new to Reverse transcription. I have worked with cDNA before but dealing with RNA extraction and reverse transcription is new for me so bare that in mind. I'm studying the...
27 December 2016 738 2 View
I am using 16S rRNA sequence for bacterial phylogeny construction.
24 December 2016 6,822 8 View
I have homologous sequences with more than 80% identity. I wonder these genes might have undergone gene duplication which resulted in their different activity (function), i.e. they are paralogs. I...
19 December 2016 4,031 11 View
Would there be any potential issues that anyone could see arising by using the RACE ready cDNA for my regular two gene specific primer PCR.
19 December 2016 8,226 1 View
I have a data set from a metatranscriptmic sample and I want to recover the 28s ribosomal sequence of one organisms present in sample. The sample had a a ribosome-depletion step before sequencing,...
18 December 2016 986 3 View
anybody know the reference or housekeeping gene of coconut hispine beetle brontispa longissima for qPCR? thanks
16 December 2016 3,427 1 View
I was following the tutorial/methods. I realized that the pipeline was applied on single-end sequencing data. I wonder if anyone has done it on paired-end data especially on the step of converted...
15 December 2016 3,674 1 View
I want to search any possible synteny of genes in both coffee and tomato genomes. I have genes sequenced and looking for if these genes maintained the same organization in tomato genome as they...
14 December 2016 696 1 View
I'm trying to figure out the best sequencing service to use in terms of price, service, quality of results, ease of sample collection, etc. What should I be considering when I select a sequencing...
13 December 2016 2,757 3 View
I am looking for a reference about the error rate in pacbio sequencing machines and how it has evolved in the recent years. I'm also interested in learning about it's randomness and how errors can...
12 December 2016 3,126 2 View
Hi everyone, I started a MiSeq run today and everything checked off as fine and I started the run. It finished in less than an hour and said "sequencing complete." However, there was not one...
12 December 2016 5,997 3 View