Summary of lectures of the mastercourse Molecular Biology of the Cell - PART I: lectures of Michel Haring and Maike Stam.
Master Biomedical Sciences - Medical Biology, University of Amsterdam
Summary Molecular Biology of the Cell 2 (book) (WBFA007-04)
Summary Regenerative Medicine
All for this textbook (62)
Written for
Universiteit van Amsterdam (UvA)
Biomedische wetenschappen
Molecular Biology of the Cell
All documents for this subject (10)
1
review
By: polhaas_95 • 6 year ago
Seller
Follow
ajvv
Reviews received
Content preview
Molecular Biology of the Cell – PART I
Michel Haring – Lecture 1: Molecular Biology: The Basics (DNA sequencing & PCR)
DNA > RNA > protein > metabolite > phenotype
DNA:
- DNA replication is semi-conservative > every new double stranded molecule consists of one old
and one new strand (left picture)
- DNA synthesis = process of the formation of phosphodiester bonds while hydrolyzing the
matching dNTP molecule (right picture)
- DNA polymerase synthesizes DNA from a double stranded “primer”
- Mistakes in DNA synthesis (normally 1 error in 105 nucleotides) can be restored through:
, • Proof-reading activity of the DNA polymerase complex (1 error per 107 nucleotides left)
• Strand-directed mismatch repair (1 error in 1010 nucleotides left)
• This does not happen in bacteria
- Chemical changes in DNA bases can cause mutation in the DNA
• Depurination = A or G deletion
• Deamination = deletion of amine group: C>T; A>G; G stops replication
• During replication: no (deletion) or any base pair will be incorporated
• Always one strand changed and one strand unchanged
• Changes can be inherited
,Sequencing:
- Dideoxy sequencing (Sanger):
• DNA synthesis while incorporating chain terminators in separate reactions for the bases
A, C, G and T
• The chain terminator (dideoxynucleotide) lacks the 3’ OH necessary for strand
extension
• DNA synthesis will stop when a dideoxynucleotide is incorporated
• Run fragments on a gel and read sequence (manually or automated using a laser and
fluorescence)
- Shotgun sequencing:
• Randomly fragmented genomic DNA (BAC library) > sequencing > assembly by
comparison of overlapping sequences
, • Contigs: assembly of smaller DNA sequences into one continuous strand
• Works well with small genomes that do not have repetitive DNA
- Pyrosequencing:
• Incorporation of a dNTP molecule supplies PPi > converted to ATP > consumed by
luciferase to yield one flash of light
• Also difficult to read repetitive sequences > higher signal when more of the same base
pairs follow up each other
Base order: ATTCCTCGG
- Ion torrent sequencing:
• H+ is released when base pair is built is > measure pH
The benefits of buying summaries with Stuvia:
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
You can quickly pay through credit card or Stuvia-credit for the summaries. There is no membership needed.
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 ajvv. Stuvia facilitates payment to the seller.
Will I be stuck with a subscription?
No, you only buy these notes for $4.81. You're not tied to anything after your purchase.
