100% satisfaction guarantee Immediately available after payment Both online and in PDF No strings attached
logo-home
Samenvatting Methoden in het Biomedisch Onderzoek 3 $6.43   Add to cart

Summary

Samenvatting Methoden in het Biomedisch Onderzoek 3

2 reviews
 315 views  23 purchases
  • Course
  • Institution

een samenvatting van alle hoorcolleges en notities. Door dit document te leren haalde ik in eerste zit een 19/20.

Preview 4 out of 70  pages

  • December 30, 2021
  • 70
  • 2021/2022
  • Summary

2  reviews

review-writer-avatar

By: aaronverbeek • 10 months ago

review-writer-avatar

By: astridhindryckxprive • 1 year ago

avatar-seller
H1: DNA


1. DNA sequentiebepaling
= volgorde van basen in DNA aflezen
a. Toepassingen
De novo genoom sequentiebepaling
- Deels of volledig
- Onbekend genoom van nieuwe organismen sequencen
Resequencing
- Versus referentiegenoom
- Mutaties zoeken
- SNPs zoeken
- Diagnose, farmacogenetica, NIPT…
Aantal DNA/ RNA moleculen tellen
b. 1e generatie
Sanger sequencing




1 primer => 1 streng per keer aflezen
2 aparte reactie = FW en RV primer
 betrouwbaarheid verhogen
- korte lengte
- moeite met herhalingen
- lage trouhhput, hoge kosten
Vroeger: 4 versch reacties met 1 ddNTP – scheiden op PAA
gel – aflezen
Nu: fluo gelabelde ddNTPs – scheiding via capillaire
gelelektroforese – aflezen door laser en sensor –
elektroferogram
Phred score => kwaliteitscore
- Q = -10*log(P)
- P = probaliteit van foutieve base call
- Hoge Q = goed
Insertie/ deletie => gemengd signaal > bulk sequencing


1

, Referentiesequentie Human genome project
mens maken Hierarchical shotgun sequencing
Sanger sequencing
Nature 2001

Human Genome sequencing by Celera
Whole genome shotgun sequencing
Sanger sequencing
Geen klonering => mapping moeilijk
Science 2001
Technologische ontwikkeling sequencing
- Automatisering
- Aanpak verandert
Ontwikkeling bio-informatica
Visie op data = open science
Biologische kennis
- Sequentie van genen
- Inzicht evolutie en conservatie
- Genetische verschillen tussen mensen
Nieuwe uitdagingen
- Volledige sequentie
- Meerdere species
- Individuele humane sequenties
- Persoonlijke genoomsequentie van iedereen
Precision medicine
- Kennis DNA en SNPs => diagnose, preventie…
- Farmacogenomics
= bepaald waarom mensen slecht reageren op drug
- Therapie op maat
- Ethische aspecten

c. 2e generatie: short read NGS
= massief parallel sequencen van geamplificeerde DNA moleculen
+ hoge throughput, lage kost
+ massale sequentiebepaling
+ snel, accuraat
+ weinig input nodig
- nog kortere readlengte
- veel fouten


2

,DOEL: 1 humaan genoom op 1 toestel op 1 dag
3 pijlers
1) Parallele detectie
- Clusters = kopieën van een template
- Klonale in vitro amplificatie van template
- Multiplexing van meerere clusters
2) Miniaturisatie
3) Integratie
= directe detectie

STAP 1: Moet complex zijn
AANMAAK DNA  Genoeg versch sequenties hebben
LIBRARY Stel: tekort => bacteriofaag DNA erbij & later weghalen




3

, Types
- WGS
- WES/ taregeted
- Amplicon sequencing
SOMS: tagmentatie & adaptorligatie samen
- Illumina Nextera kit
- Transposase: knippen & ligeren
- PCR cycli => extra barcodes en adaptoren toev
STAP 2: = in parallel verschillende templates kopiëren
KLONALE IN Solid phase bridge amplification
VITRO
AMPLIFICATIE




Emulsie




Rolling circle amplificatie in oplossing




STAP 3: Sequencing by synthesis
SEQUENCING = synthese streng complementair aan template
Nadeel: fouten bij homopolymeren & indels




4

The benefits of buying summaries with Stuvia:

Guaranteed quality through customer reviews

Guaranteed quality through customer reviews

Stuvia customers have reviewed more than 700,000 summaries. This how you know that you are buying the best documents.

Quick and easy check-out

Quick and easy check-out

You can quickly pay through credit card or Stuvia-credit for the summaries. There is no membership needed.

Focus on what matters

Focus on what matters

Your fellow students write the study notes themselves, which is why the documents are always reliable and up-to-date. This ensures you quickly get to the core!

Frequently asked questions

What do I get when I buy this document?

You get a PDF, available immediately after your purchase. The purchased document is accessible anytime, anywhere and indefinitely through your profile.

Satisfaction guarantee: how does it work?

Our satisfaction guarantee ensures that you always find a study document that suits you well. You fill out a form, and our customer service team takes care of the rest.

Who am I buying these notes from?

Stuvia is a marketplace, so you are not buying this document from us, but from seller JenteDG. Stuvia facilitates payment to the seller.

Will I be stuck with a subscription?

No, you only buy these notes for $6.43. You're not tied to anything after your purchase.

Can Stuvia be trusted?

4.6 stars on Google & Trustpilot (+1000 reviews)

70055 documents were sold in the last 30 days

Founded in 2010, the go-to place to buy study notes for 14 years now

Start selling
$6.43  23x  sold
  • (2)
  Add to cart