Plant DNA extraction - CTAB Method

 

Plant DNA extraction - CTAB Method 

An extraction of DNA from plant samples

using c tab method

100 ml of setup lysis purple can be

prepared using this description

after collection of plant samples i have

to measure

200 milligram

and grind it with liquid nitrogen

then transfer all ground

plant samples into an

append of tube and

add 700 microliters of

sit up lysis buffer

vortex the mixture then

incubate at 65 degrees Celsius

for 20 minutes in a water bath

after incubation centrifuge

samples at 10 000 rpm

for 10 minutes

after centrifuge

transfer supernatant to a clean

tube and add

an equal volume of chloroform

and isoamyl alcohol

after addition of chloroform and iso-mine

alcohol

mix it well using vortex

and centrifuge at 10 000

rpm for 10 minutes

after centrifuge you will see

an aqueous upper layer

and an organic bottom layer

now transfer

the aqueous layer

to a clean tube

while transferring the quest layer

ensure that you are not disturbing

the bottom layer

to this add

600 microliters of ice cold ethanol

and 150 microliters of

sodium chloride solution

now mix the solution and

centrifuge at 13 000

rpm for 10 minutes

after centrifuge you could see

DNA pellet at the bottom of the tube

now decant supernatant

and wash the pellet with 600 microliters

of

70 percent ethanol

after addition of ethanol

mix it then centrifuge

at 10 000 rpm for 5 minutes

after centrifuge decant

ethanol and air dry

the pellet while pouring out ethanol

ensure that palette stayed

at the bottom of a conductive

air dry the pellet

add 50 microliter of

tea buffer to the pellet

and resuspend it

after store the samples at 4 degree

Celsius for further analysis.

 

 

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