Advanced techniques in molecular biology Flashcards Preview

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Flashcards in Advanced techniques in molecular biology Deck (11)
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1
Q

Steps of molecular phylogenetics

A
  1. alignment: building the data model and extracting a dataset
  2. determining the substitution model
  3. consider sequence variation
  4. tree building
  5. tree evaluation
2
Q

Methods to manipulate gene expression

A
  • RNAi
  • over expression
3
Q

Methods used for local studies

A
  • IHC
  • quantum dots
4
Q

Methods used for functional studies

A
  • co-immunoprecipitation
  • proximity ligation assay (PLA)
  • FRET
  • FRAP (fluorescence recovery after photobleaching)
5
Q

non-coding RNA (ncRNA)-associated gene silencing can be done by

A
  • chromatin remodelling
  • transcriptional regulation
  • post-transcriptional regulation
6
Q

Microscopy methods

A
  • fluorescence microscopy
  • confocal m.
  • electron m.
  • atomic force m.
7
Q

Name some basic molecular biology techniques

A
  • DNA cloning
  • PCR
  • gel electrophoresis
  • molecular hybridisation
  • mutagenesis
  • DNA and protein arrays
8
Q

Steps of protein modeling / molecular modeling

A
  1. identify related proteins with well-known 3D structures -
  2. align the related 3D structures with the target sequence and determine those structures that will be used as a template -
  3. contruct a model for the target sequence based on its alignment with the template structure(s) -
  4. evaluate the model against a variety of criteria to determine if it is satisfactory
9
Q

What detection techniques can be used in stainings?

A
  • Fluorophore or fluorochrome
  • DAB: 3,3’-diaminobenzidine tetrahydrochlorind
  • AEC: aminoethly carbazole
  • AP: alkaline phosphatase
10
Q

Name some advantages of quantum dots over conventional fluorophores.

A
  • They are more stable
  • they have narrow spectra
  • they shine brighter
  • they last longer
  • are often less toxic
  • specific organic molecules can be attached to them
11
Q

It is proposed that histone modifications affect chromosome functions. What are the two mechanisms that are suggested to do this?

A
  1. modifications may alter the electrostatic charge of the histone resulting in a structural change in histones or their binding to DNA.
  2. the modifications are binding sites for protein recognition modules that recognise acetylated or methylated amino acids