Illumina Sequencing Overview: Library Prep to Data Analysis | Webinar

Illumina Sequencing Overview: Library Prep to Data Analysis | Webinar | Ambry Genetics

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Күн бұрын

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Scott Westenberger, Staff FAS with Illumina, will present an introduction to the Illumina Sequencing Workflow for Next Generation Sequencing. This presentation will cover the library preparation construct needed for sequencing on Illumina platforms, and the biochemical processes of cluster generation and sequencing by synthesis that are automatically performed by Illumina sequencing devices. This will provide a general overview of the process which applies to any assay across all different sequencing instruments, providing some comparison and contrast between different chemistry versions and instrument engineering developments.
Presented By:
Scott Westenberger, PhD | Presenter
Staff Clinical Field Applications Scientist, Illumina
Staff Field Applications Scientist with over 10 years in biotech industry supporting next generation sequencing and microarray devices, and over 20 years in molecular biology and genetics research. Specialist in Clinical Oncology, focused on supporting CLIA labs running NGS IVDs and Lab Developed Tests and Pharma and CROs engaged in translational research, clinical trials and companion diagnostics. Ph.D from UCLA in 2006 in Microbiology, Immunology and Molecular Genetics. Post-doc at The Scripps Research Institute in San Diego focused on parasite genomics and novel antimalarial drug screening.
Tara Namey, MS, LCGC | Moderator
Sr. Genomic Science Liaison, Ambry Genetics
Tara Namey joined Ambry Genetics in 2016 as an Oncology Genetic Specialist. Prior to joining Ambry, Tara was employed for 16 years at Lehigh Valley Health Network during which time she played an integral role in the development and growth of the Greg and Lorraine Harper Cancer Genetics Program. During her time there, Tara served as both Manager and Senior Genetic Counselor. Tara received her Bachelors of Science degree in Biology/Pre-Med from the University of Scranton. She earned her Masters of Science degree in Genetic Counseling from California State University, Northridge and is board-certified by the American Board of Genetic Counseling.

Пікірлер: 36

@nostalgia545 3 жыл бұрын

It’s so frustrating having to look up resources to learn something because the people who are suppose to teach/train you just put a picture on a slide and expect you to just know from a picture. This is the best explanation I could find also

@zeinabaltoufaili5791 3 жыл бұрын

Best explanation of Sequencing by Synthesis that I could find! Thank you

@davidgangitano7427 2 жыл бұрын

Excellent presentation and very clear about a technically sophisticated topic, Dr Westenberger!!!! Beste, Dr. G.

@Happy_Salchicha 3 жыл бұрын

Thank you, finally a well-done explanation! Very helpful!

@yahyasepahi1498 3 жыл бұрын

Thank you so much. It was a very useful video.

@igoruporov1057 2 жыл бұрын

Very nice presentation! Clear and short. I'll recommend it to my students to learn most popular platform of NGS and excellent english. Thank you very much.

@sleepyowl910 2 жыл бұрын

Excellent English? Just curious about your criteria and what qualifies one to make such judgements...

@yudhkaew6387 2 жыл бұрын

It's very new and difficult for me. But after watching this video, I'm clearly understand. Thank you very much.

@MichaelRehman 3 жыл бұрын

Fantastic presentation

@user-yd2my9vx2g 3 жыл бұрын

very helpful, thanks

@ritadecassiacavaglierimede9357 3 жыл бұрын

I loved. It was very helpful. Thank you so much.

@Imtiaz_99_1 3 жыл бұрын

You well come

@zc8198 3 ай бұрын

Fantastic! Thanks a lot!

@pietgodaard4610 3 жыл бұрын

Need miseq. Arrived here for lore. Satisfied

@rsggho2532 3 жыл бұрын

On 31:46 why are the colours that corresponds to the bases different ?

@mounikareddymouni7607 3 жыл бұрын

@usmanasghar1127 3 жыл бұрын

Great video. Can we get PPT of this lecture.

@mikewheeler1712 Жыл бұрын

how is cluster density quantified? I know it is given in units of K/mm2. How are the clusters detected initially before SBS starts?

@saabitrishrestha2811 2 жыл бұрын

Thank you. Is it that we must always use barcoded primers (instead normal primers) in PCR for Illumina NGS?

@joshua43214 2 жыл бұрын

Yes. This is a complex subject, and library prep requires a very competent tech. If you do not have a person well versed in molecular technique (and it sounds like you do not), then pay to have the library made by the people doing the sequencing. Our core charges a discounted rate of $250.00 per sample for in house labs. The cost alone should let you know this is not a trivial thing that a general lab tech should do (the adapters are very cheap).

@leifang4725 3 жыл бұрын

Back to the second question asked in the video, so if there are 5 or more Gs appeared in the first 5 images, the system won't tell if there is a missing spot for cluster or a G for the 6th read, right?

@eunicegeo7738 3 жыл бұрын

My question as well, while wondering at the same time, what are the chances of this happening (especially in WES)

@joshua43214 2 жыл бұрын

In practice it is actually 2 G's that will muck things up. Discovered this the hard way when using Nextera CD libraries and switching from HiSeq to NovaSeq

@wormball 3 жыл бұрын

How do you attach different adapters to either end of the fragment?

@codieanneedwards 2 жыл бұрын

In the kits you will have an adaptor Ligation step. Right now I’m using the KAPA hyper Prep kit which has this step. The ligation buffer helps attach the desired adaptors to the ends of our fragmented dna

@willie0527 Жыл бұрын

Does anyone know how only the reverse strands are removed from flow cell?

@merlinlautner1332 3 ай бұрын

I would assume with a specific restrictionenzyme

@Bob_Adkins 2 жыл бұрын

This is for bacteria only, correct? Is there a similar library and detection process for viruses?

@wormball 3 жыл бұрын

Why not place index sequence after the primer and sequence it at once?

@ananyagupta4824 Жыл бұрын

It’s because then you wouldn’t be able to tell the difference between the sequence insert and the index

@ds5375 2 жыл бұрын

Oh man - no dice. Anybody else here for the data analysis only? - Data Analysis is NOT discussed in this otherwise great seminar :)

@withramya 3 жыл бұрын

The second question was not answered. The question was "How will the system differentiate a deletion in the sequence vs G calling (since both will look blank)?" and not "How to differentiate between lack of signal and G"- which is what he answered.

@wormball 3 жыл бұрын

the deletion will not result in pause in sequencing