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

[@nostalgia545]

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]

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

[@davidgangitano7427]

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

[@igoruporov1057]

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]

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

[@Happy_Salchicha]

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

[@yudhkaew6387]

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

[@mikewheeler1712]

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

[@willie0527]

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

[@merlinlautner1332]

I would assume with a specific restrictionenzyme

[@leifang4725]

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]

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

[@joshua43214]

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

[@saabitrishrestha2811]

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

[@joshua43214]

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).

[@rsggho2532]

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

[@MichaelRehman]

Fantastic presentation

[@wormball]

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

[@codieanneedwards]

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

[@Bob_Adkins]

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

[@zc8198]

Fantastic! Thanks a lot!

[@usmanasghar1127]

Great video. Can we get PPT of this lecture.

[@ritadecassiacavaglierimede9357]

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

[@Al-Imtiaz]

You well come

[@pietgodaard4610]

Need miseq. Arrived here for lore. Satisfied

[@wormball]

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

[@ananyagupta4824]

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

[@withramya]

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]

the deletion will not result in pause in sequencing

[@yahyasepahi1498]

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

[@ds5375]

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

[@ervinshishmani745]

[@侯雨辰-t2x]

very helpful, thanks

[@mounikareddymouni7607]

Tq

[@noorulali1184]

Illumina tech is so old and complicated. Yeast literally does sequencing by synthesis on nanograms of sugar.

[@VMVarga-ji1vx]

ниче не поняла пожно па русски пж