DNA extraction methods Flashcards
a procedure used to isolate DNA from the nucleus of cells
dna extraction
dna extraction was done first by who?
friedrich miescher
lysis of cell and the nucleus to extract DNA to the buffer
cell dissolution
removing the impurities and proteins from the sample
precipitation
final stage and to get a completely pure DNA sample
purification
basically the isolation of nucleic acid material
dna extraction
method is based on typical chromatographic methods
chromatography-based extraction method
DNA molecules and other simple and complex molecules in a COMPLEX MATRIX can be separated based on net molecular charge or molecular size
chromatography-based extraction method
used for cell lysates
chromatography-based extraction method
the dna sample is mixed with cesium chloride
etbr-csci gradient centrifugation method
how many hours is dna to be ultra-centrifuged?
10 hours
acts as intercalation agent
ethidium bromide
sample is suspended in an alkaline solution containing NaOH and SDS detegent
alkaline extraction
meaning of sds
sodium dodecyl sulfate
added to neutralize the solution
potassium acetate
selective binding of the negatively charged DNA with the silica
surface covered with positively charged ions
silica extraction
used for dna extract
silica extraction
The DNA-containing sample is added with lysis buffer, SDS and proteinase K
ctab extraction
the initial step involves liberation of DNA from a cell using SDS
phenol-chloroform method
main reagent for phenol-chloroform method
phenol-chloroform-isoamyl alcohol
used to preserve DNA from being degraded by phenol
chloroform
a serine protease from the fungus Engyodontium album
proteinase k
can be used for crude cell lysates
salting out method
The sample is first mixed with SDS, proteinase K and lysis buffer
silica column-based dna extraction method
ideal for automation and require little equipment to perform as it does not depend on centrifugation
magnetic beads
Prior of using this, DNA can be washed from the material
cellulose-based paper
impregnated with buffers, detergents, and chelating agents that facilitate the
extraction of DNA
cellulose-based paper
what material is the chelex resin
styrene divinylbenzene
material which chelates metal ions that act as cofactors for
DNases
chelex resin
can be used for tissue samples
chelex-100 extraction
appropriate for plant cell samples
filter paper-based dna extraction method
disc used in filter paper-bases dna extraction method
whatman filter paper
