2) Next Generation Sequencing (NGS) - Sample Preparation

What is covered in this video:

For more information on this topic, please visit:

Want to learn about RNA Sequencing? Watch our FREE webinar, "The Beginner's Guide to RNA-Seq"

Watch the other videos in this series on NGS:

Check out our other video series:

Connect with us on our social media pages to stay up to date with the latest scientific discoveries:

[ToonTales]

1. Whole Genome Seq (WGS) 1:20
1) TruSeq PCR-free Library Prep Kit 1:33
2) TruSeq Nano DNA Library Prep Kit 3:07
3) Nextera DNA Library Prep Kit 3:55
4) Nextera DNA XT Library Prep Kit 5:15

2. Exome Seq 5:35
1) Nextera Rapid Capture Exome Kit 6:14
2) Nextera Rapid Capture Expanded Exome Kit 7:40

3. RNA Seq 8:05
1) TruSeq Stranded Total RNA Kit 8:24
2) TruSeq Stranded mRNA Kit 10:04
3) TruSeq Small RNA Kit 10:24

4. Methylation Seq 11:15
1) TruSeq DNA Methylation Kit

[jlowry]

The quality of this video is fantastic! Well done.

[dufo]

tip: the critical 3' adenylation of the fragments before ligation (dA tailing) is mediated by special enzymes, like the Ecoli DNA pol1 klenow fragment!

[dufo]

That was an excellent and very illuminating video, thank you
Although you have to watch it several times to fully understand it!

[TheCarbon]

Great concise video, great information and way of presentation

[huynamnguyen]

Thank for your interesting videos!

Hi, I have a few questions that need urgent answers:
1/ How flourescent label in the blocking group are removed from the nucleotide?
2/ How to wash away free nucleotide?
3/ Why do we have to sequece 2 strain of DNA?
4/ What is the function of index 1 and 2 in adapter attached to DNA?
Thanks you once again.

[sharduldhole]

Very informative video! I have a few questions,
1. In Nextera Rapid Capture Exome Kit, How can we design oligos that are complementary to the exome without knowing the sequence of the exomes?
2. How the methylated and unmethylated cytosines that are converted to cytosine and uracil respectively are distinguished further during the sequencing step?

[thanhtranhuynh]

Thank for your interesting videos!
I have questions in RNA sequencing.
1/Why do we need to use UTP nucleotide instead of TTP nucleotide?
2/ After adapter ligation, we use enzyme to remove U nucleotide and my question is that Can we use DNA fragments which possess U nucleotide after deleting U nucleotide?
Thanks you once again.

[alicepat]

I really like your videos, just a note you spelled bioanalyzer wrong in a couple places(3:02, 10:04, and 11:10)

[believeinyourself]

Kindly suggest some good book name to study ngs in detail

[dariocosemans]

What about fragments where the same adapter binds at each side? How do you make sure the fragments have two different adapters on each side?

[yinjudy]

Great video! One simple question about the adaptor ligation (after adding A on 3’ of the blunt fragments), if I understand it correctly, the two ends of DNA fragments are supposed to link with different adapters right (the orange and green colour)? As both of them have the same 3'A, how to ensure that?
I think fragments linked with the same adaptors will be produced at the same time right (orange and orange, green and green)? Do we do something specific to prevent this? Or we filter them out afterwards using the Bioanalyzer.

[biotechingenio]

Excellent video.I have a question about RNA-sequencing...at the point that you dismiss rRNA from your total RNA with magnetic beads (Ampure XP beads, maybe), how does this magnetic beads "recognize" the rRNA instead the other fragments of total RNA?
I'm asking this because in my case I need that rRNA fragments to identify bacteria by 16S rRNA and indeed...I used Ampure XP beads in order to wash away that useless material (total RNA) and I don't understand completely how they work.Thanks for your attention.

[martinmazukiewicz]

Important question for me. It's about the first Sars-Cov-2 ("WH Human 1", as the Chineses called it at that time. From the BALF taken from a patient, how could they create adaptors to catch the virus fragments as they didn't know its sequence yet ? They could't have primers. How the adaptors to catch an unknown RNA fragment can be built ? The only solution i can imagine : create four different primers with on each one one of the four single nucleotide (A, T, C, G). But is one nucléotique enough to fix the fragment ? Help !!! Thanks

[mysticalgirl]

very informative video, thank you :) How would you use NGS to identify Amyotrophic lateral sclerosis (ALS) with focus on RNA binding proteins TDP43, FUS, TAF15? What would be the approach?

[barbarad.]

which adapter ligates to the bead? the barcode-adapter or the P1-Adapter?

[raedahassan]

Background music is really noisy despite the video very useful