Useful timestamps: 0:00 Introduction 2:28 A Brief History of Genetics 4:34 Sanger Sequencing vs. Illumina Sequencing 8:40 Intro to Next Generation Sequencing 15:04 Important Considerations for Whole Genome Sequencing 17:04 Understanding the Workflow 27:00 Other Applications: Plasmid Verification, mtDNA-Seq 28:35 From the Human Genome Project to Today 30:14 Additional Resources and Conclusion
This is an extraordinary and helpful webinar! It is by far the best I've seen on NGS and WGS! And I like the way the two presenters switch with their presentations! Well done to the team! The next content I would love to see (I hope I'm speaking for many viewers) is some insight into the analysis of the sequence data. Thank you!
Hi Ahmed, thank you for watching our webinar and leaving such a nice comment! Please let us know if you have any questions about the material and we will try our best to assist you!
Very helpful for a novice like me. I love your videos explaining the science so I know what I need, and your comprehensive services. I hope to get my project off the ground with your help soon.
Very helpful video, thank you. You talked about mitochondrial DNA at the end of the video. But is the mtDNA sequenced along with the nuclear DNA during whole genome sequencing or do you have to make another run ?
What strand does the fluorescent synthesis strand complement to? The reverse strand??? Once the fluorescent base is read do we have to infer the complement nucleotide to get the original template sequence?
This depends on which library prep kit is being used. For example, for Illumina's TruSeq mRNA Stranded kit, Read 1 corresponds to the reverse or antisense strand, while Read 2 corresponds to the forward or sense strand. In contrast, a library prepared with Illumina's Nextera Technology won't be stranded or directional. Other commercial kits may vary and it is best to check with the kit supplier for more information on their particular kit.
Lifestyle inherited structures can be modified. GENE DNA mutations can define systems functionality . SEQUENCING gene nucleotide Advantage’s could be the biological answers to creating a new recombinant for Corona . DNA platforms unveil the WHOLE genome that needs to be read. THANKS for the CME review
Hii like if we want to sequence whole genome of human like 50, having covid 19 symptoms half of them and half of them are asymtomatic. Which seq technique is good for them
Hello Aniella, This depends on the goal of your study. For example, Whole Genome Sequencing looks at the entire DNA genome, while Whole Exome Sequencing focuses on sequencing the exons only (i.e. protein coding regions). In contrast, RNA sequencing looks at RNA transcribed from DNA, some of which, but not all, derives from the exonic regions. For more information on the different sequencing techniques and its applications, please visit our Knowledge Base at: info.abmgood.com/next-generation-sequencing-ngs-experimental-design
19:30 - The goal of library prep is to create clusters of DNA instead of free floating DNA fragments, and Bridge PCR allows for cluster formation to occur. During library prep, two types of adapters are added to the DNA fragment, one adapter on each side of the fragment. Two types of oligos are also fixed on the flow cell (one forward and one reverse), with each oligo on the flow cell being complementary to each adapter. During amplification, the ssDNA fragments will bend or bridge over to reach nearby matching oligos on the flow cell, allowing for the amplified DNA to be in clusters. With normal PCR, the oligos are free floating in solution in the tube and do not allow for clusters to form. After amplification, the bridges are in the form of dsDNA. A denaturation step then occurs by increasing the temperature and dsDNA becomes ssDNA. The denaturation step leaves single stranded templates that are anchored to the substrate, allowing for the next round of amplification to occur, and eventually resulting in the flow cell looking like step 5.
But what define the "sequence" ? Is it in 3D, 2D or time space? in other words, if one read AGT, what made that special order A-G-T? Why cannot it be read out as T-A-G ?
Because that is the order that the DNA polymerase or RNA polymerase reads the sequence, also known as the "sense" sequence. You don't read a book backwards, you start from page 1 and read to the last page.
Hello Michael, To determine the sequence, all four types of nucleotides are added during sequencing. After nucleotide incorporation, the remaining non-incorporated nucleotides are washed away. Next, a camera takes images of the fluorescently labeled nucleotides (i.e. fluorescent signal is read at each DNA cluster and recorded). The fluorescent molecule, along with the terminal 3' blocker, is then chemically removed from the DNA and washed away, allowing for the next cycle to begin. This process is repeated until the sequencing reaction is complete and allows for only a single nucleotide to be captured per cycle.
I had to listen to it over and over again the figure out that she was saying "throughput" around 5:00 Maybe clean up the narrative on these videos, people shouldn't have to struggle to understand.
Hi there! Were you interested in chloroplast genome sequencing? If so, please contact our NGS Specialists at ngs@abmgood.com and they will be happy to assist you with your research.
the voice of the speaker (in the start) , her tone of voice and her pronunciation is really very annoying. addition of subtitles would be very helpful.
We'll take your feedback into account when we are working on our future videos! Meanwhile, you can try turning on the English (auto-generated) captions since we found them to be fairly accurate.
Useful timestamps:
0:00 Introduction
2:28 A Brief History of Genetics
4:34 Sanger Sequencing vs. Illumina Sequencing
8:40 Intro to Next Generation Sequencing
15:04 Important Considerations for Whole Genome Sequencing
17:04 Understanding the Workflow
27:00 Other Applications: Plasmid Verification, mtDNA-Seq
28:35 From the Human Genome Project to Today
30:14 Additional Resources and Conclusion
Whole genome means whole DNA or a gene of a DNA , part of gene is amplicon????
This is an extraordinary and helpful webinar! It is by far the best I've seen on NGS and WGS! And I like the way the two presenters switch with their presentations! Well done to the team! The next content I would love to see (I hope I'm speaking for many viewers) is some insight into the analysis of the sequence data. Thank you!
Hi Ahmed, thank you for watching our webinar and leaving such a nice comment! Please let us know if you have any questions about the material and we will try our best to assist you!
Very helpful for a novice like me. I love your videos explaining the science so I know what I need, and your comprehensive services. I hope to get my project off the ground with your help soon.
Thanks so much for watching! ;) Glad you found the video helpful!
Very helpful video, thank you. You talked about mitochondrial DNA at the end of the video. But is the mtDNA sequenced along with the nuclear DNA during whole genome sequencing or do you have to make another run ?
Welldone very helpful webinar regarding WGS, please arrange webinar on Multi locus sequence typing
Very clear!!!!!!!!!!!!!
Thanks for watching!
any video on new era sequencing ?
What strand does the fluorescent synthesis strand complement to? The reverse strand??? Once the fluorescent base is read do we have to infer the complement nucleotide to get the original template sequence?
This depends on which library prep kit is being used. For example, for Illumina's TruSeq mRNA Stranded kit, Read 1 corresponds to the reverse or antisense strand, while Read 2 corresponds to the forward or sense strand. In contrast, a library prepared with Illumina's Nextera Technology won't be stranded or directional. Other commercial kits may vary and it is best to check with the kit supplier for more information on their particular kit.
Can you add subtitles to the video？This will make it clearer and easier to watch.
Hi Bo, if you click the "CC" button on the bottom right of the video player, there is an option to turn on subtitles for the video.
I just subscribed
Lifestyle inherited structures can be modified. GENE DNA mutations can define systems functionality . SEQUENCING gene nucleotide Advantage’s could be the biological answers to creating a new recombinant for Corona . DNA platforms unveil the WHOLE genome that needs to be read. THANKS for the CME review
Hii like if we want to sequence whole genome of human like 50, having covid 19 symptoms half of them and half of them are asymtomatic. Which seq technique is good for them
Hello Aniella,
This depends on the goal of your study. For example, Whole Genome Sequencing looks at the entire DNA genome, while Whole Exome Sequencing focuses on sequencing the exons only (i.e. protein coding regions). In contrast, RNA sequencing looks at RNA transcribed from DNA, some of which, but not all, derives from the exonic regions. For more information on the different sequencing techniques and its applications, please visit our Knowledge Base at: info.abmgood.com/next-generation-sequencing-ngs-experimental-design
What is the point of the DNA forming bridges like this? And how are the bridges then disrupted for the chip to look as it does in fig 5?
19:30 - The goal of library prep is to create clusters of DNA instead of free floating DNA fragments, and Bridge PCR allows for cluster formation to occur.
During library prep, two types of adapters are added to the DNA fragment, one adapter on each side of the fragment. Two types of oligos are also fixed on the flow cell (one forward and one reverse), with each oligo on the flow cell being complementary to each adapter.
During amplification, the ssDNA fragments will bend or bridge over to reach nearby matching oligos on the flow cell, allowing for the amplified DNA to be in clusters. With normal PCR, the oligos are free floating in solution in the tube and do not allow for clusters to form.
After amplification, the bridges are in the form of dsDNA. A denaturation step then occurs by increasing the temperature and dsDNA becomes ssDNA. The denaturation step leaves single stranded templates that are anchored to the substrate, allowing for the next round of amplification to occur, and eventually resulting in the flow cell looking like step 5.
@@abmgood Thank you.
But what define the "sequence" ? Is it in 3D, 2D or time space? in other words, if one read AGT, what made that special order A-G-T? Why cannot it be read out as T-A-G ?
Because that is the order that the DNA polymerase or RNA polymerase reads the sequence, also known as the "sense" sequence. You don't read a book backwards, you start from page 1 and read to the last page.
Hello Michael,
To determine the sequence, all four types of nucleotides are added during sequencing. After nucleotide incorporation, the remaining non-incorporated nucleotides are washed away. Next, a camera takes images of the fluorescently labeled nucleotides (i.e. fluorescent signal is read at each DNA cluster and recorded). The fluorescent molecule, along with the terminal 3' blocker, is then chemically removed from the DNA and washed away, allowing for the next cycle to begin. This process is repeated until the sequencing reaction is complete and allows for only a single nucleotide to be captured per cycle.
TB whole genome sequencing
How does RNA contamination degrade our DNA samples?
I had to listen to it over and over again the figure out that she was saying "throughput" around 5:00
Maybe clean up the narrative on these videos, people shouldn't have to struggle to understand.
Assalamualaikum mohon ijin kami Jalasenastri Ny.eka Agus Riyanto mohon ijin bergabung'🙏
Chloroplast DNA
Hi there! Were you interested in chloroplast genome sequencing? If so, please contact our NGS Specialists at ngs@abmgood.com and they will be happy to assist you with your research.
the voice of the speaker (in the start) , her tone of voice and her pronunciation is really very annoying. addition of subtitles would be very helpful.
We'll take your feedback into account when we are working on our future videos! Meanwhile, you can try turning on the English (auto-generated) captions since we found them to be fairly accurate.
Your comment is witless, everything about the first speaker was excellent.
Wow, how racist, grow up.