bacteria transformations

[Smiley_]

how much info do we need to know?

[Snorlax]

How much info do you know?
All my textbook has on it is that when a recombinant plasmid is taken up by a bacteria cell, that's called transformation.

[Smiley_]

How much info do you know?
All my textbook has on it is that when a recombinant plasmid is taken up by a bacteria cell, that's called transformation.

yeahs that pretty much it

[Yacoubb]

Know how a plasmid can be recombined. That is, restriction enzymes are used to cut and isolate a particular gene, like the human insulin gene. The same restriction enzyme cuts out and isolates a portion of the plasmid to allow the human insulin gene to be inserted into the fragment to join permanently. This is achieved by ligation (using ligase enzymes).

Bacteria must be competent in order to take up the plasmids. That is, they must be bathed in calcium chloride solution and then be heat shocked (heated to 42^oC for 90 Seconds), in order for the bacterial cell walls to become permeable to the transformed plasmids and actually take them up. Not all the bacteria will take up the plasmids. Only the bacteria who have taken up the plasmids with the recombined DNA will be transformed.

Hope this helped

[wallah]

Just while we are discussing about Bacteria, I wanted to know if any kind of staining can kill bacteria or not?

[Yacoubb]

Just while we are discussing about Bacteria, I wanted to know if any kind of staining can kill bacteria or not?

Why would you stain bacteria? :/

[wallah]

there was a question which asked how can you find out whether the bacteria has taken up the plasmid or not (performed transformation) so I was wondering if you could stain the plasmid, as well as on the other hand you could just test it's resistance towards certain solutions. But will stating. "stain the plasmid so it is visible," get you the marks?

[Yacoubb]

there was a question which asked how can you find out whether the bacteria has taken up the plasmid or not (performed transformation) so I was wondering if you could stain the plasmid, as well as on the other hand you could just test it's resistance towards certain solutions. But will stating. "stain the plasmid so it is visible," get you the marks?

If you place a pGLO gene onto the plasmid, the bacteria that have successfully taken up the recombinant plasmids will glow in the dark. You could also test its resistance to antibiotics by culturing a bacterial colony around a ring of the antibiotic; those that can grow are clearly resistant.

Those are two ways of identifying successful transformation.