Questions tagged [dna-sequencing]

Technique(s) by which the sequence of DNA is obtained. The principles are similar for RNA-sequencing.

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Why do genetic testing companies (FTDNA,AncestryDNA,23andme) express DNA shared in centimorgans (cM) instead of in number of base pairs or in percent?

Expressing DNA shared in number of base pairs or in percent of total genome would be so much simpler and easier to understand for the customers. .
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Any examples of a group of micro-organism that continuously spans two or more species classifications?

Are there any examples of a group of micro-organisms where two different, established species are designated and these two groups meet all aspects of the definition of species (perhaps can not ...
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143 views

Ray wu DNA sequencing methodology understanding

I'm trying to understand the method which Ray Wu, developed to sequence the DNA 5' cohesive ends of a lamda phage. I'm reading these two papers written by him, but i stuck at the point where he uses ...
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Why is a screening process required in selection from a genomic DNA library?

After the process of construction we go for screening process of selecting a recombinant host. Which includes: DNA hybridisation Colony hybridisation Random primer labeling Nucleotide ...
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72 views

C to T Transition Mechanism

I am seeing C to T transitions in my sequencing data, but also some strange signatures that follow along with that. I am trying to understand the biology of what is occurring. In the data I see more C ...
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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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470 views

dNTP concentration for reverse transcription from RNA to cDNA

Does it affect the results if I use the half of dNTP amount in a reaction that it says in my protocol? I usually use 4 microlitres per reaction and this time I did a mistake and I used 2. The final ...
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42 views

Error frequency of DNA replication without proof reading activity of DNA polymerase?

Different domains of DNA polymerase contain different activity, like 5'->3' polymerisation and 3'->5' proof reading ...
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387 views

Limitations of Gibson Assembly method

Why is the Gibson assembly method limited to ~8 fragments and what can be done to raise this limit? Why is the method called "seamless" and what are the advantages compared to annealing ...
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128 views

what is CpG coverage? DNA starting material?

referring to this quote from an article about predicting methylation states: "Because of the small amounts of genomic DNA starting material per cell, single-cell methylation analyses are ...

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