what are the steps involved in biotraansformation?
Answers
Answered by
1
So, in a transformation, you are trying to get bacteria to suck up your DNA so that you can trick them into making a protein you want them to make.
The first step is to get competent cells (cells that are willing to suck up the DNA) you can do this by either buying them or making them by incubating them in a calcium chloride solution (I haven't done this part for a while so I don't remember the exact steps).
The next step is to incubate your DNA plasmid with the bacteria and then shock them into incorporating the DNA. The most common way to do this is heat shock, where you heat the sample to 42 degrees C for ~45 sec then put it on ice for a minute or two (protocols differ on the numbers).
Next you dilute the bacteria in some sort of media and shake them at 37 degrees C for an hour or two so that they can grow
The final step is to select for the bacteria who are successfully transformed. This is usually done by having a gene inserted into your original plasmid which would make the transformed bacteria immune to a certain antibiotic. You then plate the bacteria on a agar plate that has that antibiotic in it and let the bacteria grow at 37 overnight.
The bacteria that grew should be the successfully transformed bacteria.
The first step is to get competent cells (cells that are willing to suck up the DNA) you can do this by either buying them or making them by incubating them in a calcium chloride solution (I haven't done this part for a while so I don't remember the exact steps).
The next step is to incubate your DNA plasmid with the bacteria and then shock them into incorporating the DNA. The most common way to do this is heat shock, where you heat the sample to 42 degrees C for ~45 sec then put it on ice for a minute or two (protocols differ on the numbers).
Next you dilute the bacteria in some sort of media and shake them at 37 degrees C for an hour or two so that they can grow
The final step is to select for the bacteria who are successfully transformed. This is usually done by having a gene inserted into your original plasmid which would make the transformed bacteria immune to a certain antibiotic. You then plate the bacteria on a agar plate that has that antibiotic in it and let the bacteria grow at 37 overnight.
The bacteria that grew should be the successfully transformed bacteria.
paulinesimon19:
Thank you Tnrai...!
Answered by
1
I have attached a file or doc go through it.
May this will help u
May this will help u
Attachments:
thank u simran!
ur welcome
Similar questions