3 Flashcards
Tissue specific?
cDNA Library
Methods for identifying specific proteins with specific antibodies?
Immunocytochemistry and western blotting
Role of mRNA and oligo(dT) in in vitro cDNA system?
mRNA- template
oligo(dT)- primer
Methods to prove actin gene expression in liver cells?
Northern and Western blotting
Radioactive precursor to label products of PCR?
[3H] dTTP
[alpha- 32P] dCTP
Difference between dTTP and ddTTP?
Absence of -OH group on 3’ position in ddTTP
Use of SNP chips?
To identify many single nucleotide polymorphisms units DNA samples
What is GMO?
Genetically modified organisms
Use of cDNA chips?
To analyze expression of many genes in a cell
Visualize result of molecular hybridization?
Autoradiography
Subcellular fractions and order of generation?
Nuclear — mitochondrial — microsomal — ribosomal — cytosol
Cellular organisms in microsomal?
Golgi, RER, SER
Separation methods for 2D electrophoresis of proteins?
Isoelectric focussing and SDS gel electrophoresis
Charge of proteins in solution with pH lower than isoelectric point of the protein?
+
Charge of proteins at the end of isoelectric focussing?
No charge
Charge of proteins at the end of SDS Page?
-
Electrophoretic method for protein separation based on mass?
SDS page
Methods independent of mass
Affinity chromatography
Isopycnic density gradient centrifugation
Relationship between elution volume and mass?
Higher elution volume = lower mass
Ligands used to separate during affinity chromatography and what they separate?
Antigen — antibody
Hormone — receptor
Separation of poly(A)+mRNA from RNA fraction?
Oligo(dT) affinity chromatography
Type of electrophoresis in DNA sequencing?
Capillary electrophoresis
