Technical tools for the study of protein and nucleic acids
Sequencing : determination of the primary structure of an unbranched biopolymer
Production
Extraction
Purification
Purification from natural sources
De novo chemical production
Heterologous expression in a model organism
Osmotic lysis
Mechanical lysis
Detergent-based lysis
Precipitation
Dialysis
Chromatography :
Electrophoresis
Protein sequencing
DNA sequencing
Sanger sequencing : the process of selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication
NGS : production of massively parralelised short reads which overlap to some extent allowing reconstruction of the final sequence
From DNA to protein:
Illumina
Nanopore sequencing
SMRT sequencing
Amino acids analysis
Edman's degradation
Making smaller poplypeptides from big proteins
Mass spectrometry
Osmotic lysis of cell placed in a hypotonic medium
Based on osmosis = tendancy of a solvent molecules to move through a selectively permeable membrane to equalize the solute concentration 
Cells are disrupted mechanically, by bead grinding, homogeniser, French press, sonicator or freeze-thawing
Non ionic detergent (+Lysozyme)
Then : centrifugation to get rid of cellular debris
Based on solubility
Change of ionic strength of a solution can cause some protein to precipitate but not others
Use of a semi permeable membrane to separate molecules, based on their size
Based on motion of particle in fluid (often polyacrylamide or agarose) under the influence of a uniform electric field
Analyzing the deplacement of a sample in the medium allows to separate molecules based on charge, molecular weight, or shape
Size exclusion chromatography (based on size)
Technique for mixture separation based on two phases : mobile phase (=liquid carrying the dissolved mixture) will pass through solid phase
Planar chromatography
Column chromatography
Ion exchange chromatography
Principle : smaller particles can enter the beads (solid phase) and take more time than big particles that by-pass the beads
Based on charge interactions between proteins and solid phase
Affinity chromatography
Based on selective non covalent interactions between analyt and a specific molecule
Adsorbtion chromatography
Overexpress a candidate DNa to see if we obtain the same protein and if we do find the protein sequence from the DNA sequence
Ion exchange
Reverse phase
AAs go through sulfonated matrix based on their isoelectric point and are then labeled with nyhydrin to be detected quantitatively
AAs are stained with phenylisothiocyate and then run through RP HPLC
N terminal AA is selectively cleaved by phenylisothiocyate and the operation is repeated
Separation of polypeptide chains and then fragmentation of polypeptide chains using proteases
