much appreciated, helps a lot!
your company's site is prefect
Cheers,
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I'd actually recommend using fosmids, rather than BACs or cosmids, but the principle is the same.Here's how to do the calculation (for more details, download the PDF from the first link on this page.)
Determining the Approximate Number of Clones for a Complete Fosmid Library:
Using the following formula, determine the number of fosmid clones required to reasonably ensure that any given DNA sequence is contained within the library.
N = ln (1-P ) / ln (1-f )
Where P is the desired probability (expressed as a fraction); f is the proportion of the genome contained in a single clone; and N is the required number of fosmid clones.
For example, the number of clones required to ensure a 99% probability of a given DNA sequence of E. coli (genome = 4.7 Mb) being contained within a fosmid library composed of 40-kb inserts is:
N = ln (1 – 0.99) / ln (1 – [4 x 10^4 bases / 4.7 x 10^6 bases]) = –4.61 / –0.01 = 461 clones
Hope that helps.Leave a comment:
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Edit
hello all,
So I came to the conclusion that I should use a combination of cosmid and BAC libraries, as was used in the pibmed article 16372010
my issue now is how to use the following equation:
Genome coverage = (total # of clones - # of non-contributing clones) X
average insert size/haploid genome size
according to the poisson distribution, the probability of obtaining one or more clones per locus is a function of library size
in my case, I need 8fold coverage which would be 99.97%
btw, the genome im working with is 20Mb
i knw that in terms of cloning capacity, cosmids can take about 40 kb
and BAC can take up to 300kb
but I cant seem to put it all together
Common seqanswer community! help a newbie out
thank youLeave a comment:
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Capillary DNA sequencing
hello all,
So my question is: is there a particular type of library i need to construct if i am to use capillary sequencing?
This is a question for an HW of mine, but I want to emphasize that I DONT WANT you to give me the answer!
I would just like to be pointed in the right direction, in terms of the theory behind building the appropriate library
Thank you
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...Yesterday, 07:11 AM -
Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
...06-02-2026, 10:05 AM -
With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.
Introduction
Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...05-22-2026, 06:42 AM
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