1. Coverage 1:01 2. Sample Quality Control 2:15 3. Quantitative Validation 3:19 4. Qualitative Validation 4:48 1) Introduce gel into the chip and pressurize it 2) Load markers, ladders, and samples onto the chip 3) Vortex the chip 4) Load the chip onto the Bioanalyzer
Hi Khushali! Thanks for watching and posting your question! We actually have a blog post that covers this topic and explains a bit more: info.abmgood.com/blog/next-generation-sequencing-what-are-sequencing-coverage-and-coverage-mapping. Essentially, there are many factors such as sequencing errors, contaminated samples, differences in thresholds and more that may affect sequencing coverage. Hope this helps!
How can we relate the protein fragments in a cell with the DNA sequencing? If i have protein expressed under some stress in plant..how can i know the RNA sequence or DNA sequence related to the protein..
If you can isolate your protein of interest, you can use mass spectrometry for protein sequencing and identification. Once you have the sequence of the protein, you can use bioinformatics tools, such as BLAST to compare your protein sequence to databases of known proteins; this can help to identify the gene that may encode your protein of interest. Hope this helps!
I have an assignment(experiment to perform next generation sequencing on 10 patients with a healthy liver and 10 patients with a liver that has a genetic disorder and write the difference between their gene expression.and what will be the outcome of next generation sequencingDon't know where to start
Hi hiatussemilunaris, thanks for the question! NGS can indeed be used to detect aneuploidy. Simply put, you perform Whole Genome Sequencing on the sample(s), and also on reference samples with known aneuploidy, and using bioinformatics analyses, quantitatively compare the ratio of uniquely mapped reads (i.e. non-duplicate reads) to the reference samples. For example, in Down syndrome samples, you would expect to see 1.5-fold higher copy number change for chromosome 21 than with non-affected samples.
Hi Santhu, coverage is the number of times a particular nucleotide is sequenced. On the other hand, reads are referring to the output of an NGS sequencing reaction. A read is a single uninterrupted series of nucleotides representing the sequence of the template. Hope this helps!
1. Coverage 1:01
2. Sample Quality Control 2:15
3. Quantitative Validation 3:19
4. Qualitative Validation 4:48
1) Introduce gel into the chip and pressurize it
2) Load markers, ladders, and samples onto the chip
3) Vortex the chip
4) Load the chip onto the Bioanalyzer
Thanks for yet another interesting video!
Very informative video.
It would be great if you could please explain which factors affect the coverage during sequencing and also how?
Hi Khushali! Thanks for watching and posting your question! We actually have a blog post that covers this topic and explains a bit more: info.abmgood.com/blog/next-generation-sequencing-what-are-sequencing-coverage-and-coverage-mapping. Essentially, there are many factors such as sequencing errors, contaminated samples, differences in thresholds and more that may affect sequencing coverage. Hope this helps!
The link about the coverage doesn't really work for me
How can we relate the protein fragments in a cell with the DNA sequencing? If i have protein expressed under some stress in plant..how can i know the RNA sequence or DNA sequence related to the protein..
If you can isolate your protein of interest, you can use mass spectrometry for protein sequencing and identification. Once you have the sequence of the protein, you can use bioinformatics tools, such as BLAST to compare your protein sequence to databases of known proteins; this can help to identify the gene that may encode your protein of interest. Hope this helps!
I have an assignment(experiment to perform next generation sequencing on 10 patients with a healthy liver and 10 patients with a liver that has a genetic disorder and write the difference between their gene expression.and what will be the outcome of next generation sequencingDon't know where to start
How did it go 🤔
how can NGS detect aneuploidy?
Hi hiatussemilunaris, thanks for the question! NGS can indeed be used to detect aneuploidy. Simply put, you perform Whole Genome Sequencing on the sample(s), and also on reference samples with known aneuploidy, and using bioinformatics analyses, quantitatively compare the ratio of uniquely mapped reads (i.e. non-duplicate reads) to the reference samples.
For example, in Down syndrome samples, you would expect to see 1.5-fold higher copy number change for chromosome 21 than with non-affected samples.
Can you please update me what is coverage and reads?
Hi Santhu, coverage is the number of times a particular nucleotide is sequenced. On the other hand, reads are referring to the output of an NGS sequencing reaction. A read is a single uninterrupted series of nucleotides representing the sequence of the template. Hope this helps!
Applied Biological Materials - abm Thank You so much...Very clear answer....
No problem! :) Our knowledge base (link in the description) has a glossary of other useful terms as well.