Technique(s) by which the sequence of DNA is obtained. The principles are similar for [tag:rna-sequencing].

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This Sequence Data (DNA) has very few Methionin-starts. How is that possible?

I'm working on my first sequencing related project and I'm trying to find proteins with a specific PFAM ID (PF11999). The project is called "MMETSP", I searched the annotations for that ID, identified ...
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16 views

Is DNA sequencing coverage a function of sample purity?

How is coverage affected by the purity of a sample? And can coverage for a sample be affected by other things, like the library preparation or manner in which the sample was stored?
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19 views

DNA Sequencing Types & Qualities

I've been searching for a list of the types of DNA sequencing (e.g. Sanger, Next-Generation) and how prone they are to sequencing errors (None, Somewhat, Very), but I haven't been able to find ...
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mtDNA sequencing error rates

What is the method to estimate the error rate in mtDNA sequencing? E.g. we are given a profile like this: 16069T 16126C 73G 152C Is there a method, a table of known values, or any other way to say ...
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25 views

Distance between P5 and custom read 1 primer for Illumina HiSeq sequencing

I have a minigene library which is amplified with P5/P7 sites that looks like this [P7/barcode][constant region][ variable region ][18nt constant region][SP1][P5] (436nt total) We sequenced this ...
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41 views

Parameters of Variant calling analysis [closed]

What is the good or stringent parameter for variant calling. At present using the DP > 10 and Q > 30 for Variant calling. Is it ok.? Thanks
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30 views

what percent of DNA consider to be “junk”?

Does 98% of a whole human genome observe to be "junk" or 98% of each single human DNA observe to be "junk"or "non protein coding region"?
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52 views

How should “the human genome” be interpreted?

What does sequencing "the human genome" mean? Don't individuals have unique genomes?
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24 views

How does one apply Bayesian inference to quantify a read the deeper you sequence?

For NGS sequencing technology, the "deeper" you sequence given fragments, the more certain you are of what is being sequenced. This sounds like a simple application of Bayes's Rule. What is the ...
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73 views

What percentage of a Neanderthal's DNA could be in a Denisovan?

My questions arose from an online course video at 3:11-19 The professor mentioned that the Neanderthals were inbred with their cousins breeding together but the Denisovans were very diverse , with 70%...
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30 views

How do I know if the tRNA gene is under purifying selection from looking at a sequence dataset?

I'm having trouble figuring out how to tell if a tRNA gene is under purifying selection, just by looking at its sequence dataset. Such as this one:
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55 views

Perplexing by the way of estimating the shared or different genes between humans and chimps

We share 98.5% genes with chimps (it means we have 98.5% same DNA sequeces ),so there is about one percent difference .It means we can approximately differ from them by one base pair every hundred ...
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84 views

What is the another use of de Bruijn graphs in bioinformatics except DNA assembly? [closed]

I implemented own generic de Bruijn graph, which I use for DNA assembly (alphabet: A, C, G, T). I try to find purpose of de Bruijn graph for another bioinformatics problems, if some exists. I want use/...
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59 views

Does the cell have a mechanism to determine DNA sequence from protein?

Something like a reverse genetic code.
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2answers
79 views

What is the frequency distribution of each base in a DNA sequence? [closed]

Can we say that the frequency distribution of each base in a DNA sequence is equiprobable? After the negative answer; i rephrase: Is there a use case in which the frequency distribution of the ...
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2answers
41 views

Genes that exist in old Affymetrix platform but not in the newer one

I am using two gene expression datasets from an Affy U95Av2 platform and an Affy U133 Plus 2.0 platform. When I map the Affy probe names to HUGO gene names, there are thousands of genes which exist in ...
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1answer
43 views

Paired-end illumina sequencing

I'm tying to understand the mechanism of paired end illumina sequencing, however I dont't understand why index primer 1 and index primer 2 are used. paired end illumina sequencing (YouTube). I ...
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57 views

Online multiple sequence alignment with constraints

This is probably a naive question. I have accesssed the Clustal Omega online multiple sequence alignment tool at http://www.ebi.ac.uk/Tools/msa/clustalo/ for the first time, but I think I need to ...
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2answers
47 views

How exactly are gaps defined in genomics?

My understanding is that when sequencing a genome, "gaps" refer to fragments which were not sequencing in the sequencer. Is this correctly? What are there other meanings to the term "gap"?
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57 views

How do high-throughput/NGS sequencers calculate quality scores?

I am confused as to how quality scores are actually calculated by DNA sequencers like Illumina. For each base call, some quality predictor value is computed, based on various properties of the ...
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38 views

How to construct tumor phylogenetic tree?

I would like to know if anyone has tried any software that constructs tumor evolution trees where the trunks represent the common mutations and the private alterations are noted on each branch. I can ...
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99 views

Why do we encode information in DNA in binary and not in base 4?

I recently read an article about Harvard scientists encoding 700Tb of data in DNA strands. But they encoded the information in base 2, so T and G was a 1 and C and A was a 0. But why binary? Why didn'...
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2answers
70 views

Do plants have distinctive DNA genomes from each other like humans do?

Can exact same species of plant have a distinct genome from others of same exact species growing nearby or in a different place/country etc. ? Can a leaf be traced to the the exact plant based on DNA ...
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340 views

In percentage, how much is the human genome (DNA) similar to the mouse genome?

Some guy argued with me against evolution theory, and he claimed that human and mice share 98% just like human and chimpanzee. I've tried to search online for a simple and accurate answer, but I ...
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1answer
63 views

Questions on adding a protein to a DNA library [closed]

Two questions regarding finding the DNA sequence of a amino acid sequence (AA): 1) If you are able to find out the mRNA sequence of an AA, then don't you automatically know the DNA sequence? 2) ...
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2answers
79 views

How to compare Smith–Waterman algorithm implementations?

Assume that you have two implementations of the Smith–Waterman algorithm (with what ever heuristic they apply to speed up) for local sequence alignment of genomic sequences. I would like to know if ...
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1answer
49 views

Are there any major noticeable limitations to genome sequence compression methods that use reference templates?

Recently, I have been researching about big data analytics in biochemistry, and started wondering about how genome sequence compression could affect analysis. Of all the method listed on the ...
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15 views

Trouble identifying a gene given an Expressed sequence tag

I have an assignment where we need to answer several questions about an EST, however, I'm having trouble getting started. My sequence contained a Not I site (clipped that) and no poly A signal. We ...
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31 views

Determine OTU identity using Blastn full database or organism specific database?

I am seeking opinions on the best way to determine OTU identity using Blastn. Would the best way to identify an OTU be to blast the OTU to the full nr/nt Nucleotide collection or to blast your OTU to ...
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37 views

Which is the proportion of metagenomics reads that cannot be mapped in any genome?

I was wondering if anybody knows how many reads from metagenomic or metatranscriptomic data do not map to known sequences and are therefore unidentifiable? I found a figure for viromics, which seems ...
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120 views

Whole gene sequence analysis to determine source infection [closed]

Is it possible to use whole gene sequence analysis to distinguish between a common source infection and a person-person disease transmission?
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1answer
59 views

constitution of read and gene region (IGV)

I work with fastq files containing NGS reads for some human DNA regions. The reference genome is hg19. I had two fastq files (pair-ended). I generated alignment BAM files. I used "bwa" and samtools to ...
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29 views

Do eukaryotes assimilate DNA that is floating in the extracellular membrane?

Prokayotes, which replicate primarily using binary fission, don't get much genetic diversity. For this reason, they take in any genetic material they encounter, in a gambit to help them better adapt ...
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28 views

Tool for translating chomosome, index numbers to sequence information

Is there a tool which allows me to transform information in the form (chromosome number, start index, end index) to sequence information? For example, something like (6, 43770819, 43770841) should ...
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40 views

Sodium hydroxide grade in library denaturation for NGS

If the NaOH, 1N which I bought does not have the label of molecular biology grade can it be used safely for library denaturation in NGS? Thank you!
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24 views

Transgenic Plant-Gene source for introduction into host organism

Let's say the plant-gene itself is known, i.e. the exact neucleotide sequence is available in a gene bank (originally sequenced from a tree sample extracted RNA). Previous work has already successfuly ...
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52 views

new DNA sequencing technologies? [closed]

Last year I did my thesis in the field of a new method for DNA analysis. Which I think is very interesting although I cannot seem to find any new methods in this field especially the SMRT sequencing ...
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59 views

How does a test like the Natera Panorama distinguish the child's genome from the mother?

Blood tests on an expectant mother, like the Natera Panorama, are now being used regularly to screen fetuses for chromosome abnormalities. At my wife's recent prenatal visit, she wasn't really even ...
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69 views

RAD sequencing: choosing the appropriate enzyme?

I’m studying Darwin’s finches genome and I say in some articles that the researchers used restriction enzymes to cut the DNA in their double digest RAD protocol. They are using EcoRI and MseI (GAATTC ...
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Can we take advantage of nanopore sequencing systematic errors to predict secondary structure motifs?

One of the methods of single-molecule sequencing, Nanopore sequencing, is based on traversal of DNA strand through a nanopore. Nucleotide is determined by measurement of ion current (when nucleotide ...
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1answer
37 views

Is there any websites that provide the DNA sequences of food samples?

We can get many DNA sequences at NCBI websites for free. Is there any websites that provide the DNA sequences of food especially meat, for free? When I read papers regarding food authentication, ...
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95 views

Why are sequencing reads shorter than PCR products?

I have desinged and tested primers for RT-PCR, then purified the PCR products from the gels and send them to sequence at GATC (SUPREMERUN) using the forward primers. After blasting the reads to ...
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How to identify genes in Ralstonia that synthesize PHB and promote granule formation?

The compound polyhydroxybutyrate (PHB) is of considerable industrial interest as a biodegradable substitute for plastic. PHB is synthesized from glycerol by the bacterium Ralstonia eutropha. PHB ...
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37 views

Determining the Accuracy of DNA Testing

The DNA Testing firm in the below article claims that its tests were "99.99 per cent" accurate. http://www.telegraph.co.uk/news/uknews/1322077/False-DNA-test-led-father-to-reject-daughter.html ...
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37 views

Where can I find a file listing the genetic differences in humans?

I don't know much about genetics. For an arts project I'm looking for a human genome. I read that: Only about 0.1% of the genome is different among individuals, which equates to about 3 million ...
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120 views

Standard practice for generating rarefaction curves from Next Generation Sequencing data

We have a few million 18S reads from a particular environment. The reads have been clustered into Operational Taxonomic Unit (OTU), and the OTUs annotated against a reference database. To generate a ...
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79 views

Does increasing the value of k in k-fold coverage requires more memory?

Study: Here is what is k-fold coverage which is know as sequence coverage. According to this, 2-fold coverage means the whole DNA is sequenced more twice in average. Question: If I have a genome ...
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1answer
101 views

How can I generate a random DNA sequence?

I've found this paper which involves the construction of 19-bp random DNA sequences, but I don't know enough biology to understand how this method works. Could someone explain it to someone who is ...
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1answer
31 views

Gemini usage of --sample-filter

I am using gemini to query databases made from vcf files, which contain data from multiple samples. However, I need to query data only from one sample, (for which I currently use grep to filter the ...
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2answers
227 views

In Sanger sequencing, why do we resort to cloning? Why doesn't PCR suffice?

I understand that in Sanger sequencing we can clone our fragments with the help of e.g. bacteria to make multiple copies of our fragments for further analysis. I also understand cloning can be a ...