Hello everyone, I will show you the transformation of a plasmid into E.coli bacteria, which will be fed to my worms for a gene knockdown.
So, 1st I took out plasmid and competent cells from -80 °C and put them on Ice to thaw.
I added, my plasmid to the competent cells waited for 5 min and did a heat shock to open up the pores of the bacterial cell wall and plasmid goes inside and then immediately put it into ice to avoid damage.
then for 5 mins later added LB medium was without any antibiotics and incubated at 37 °C for the bacterial growth for 1hr. in the meantime filled some tubes and kept it for autoclaving.
now incubation is over and we can spread our bacterial culture 10- 30 ul and spread in on LB ampicillin plate with a glass spreader and incubate at 37 degree Celsius and the next day our bacterial colonies appear.
16 сен 2024
