How do we know which transformation is occurring with blue/white screening?

In summary, blue/white screening is a technique used to distinguish between successful and unsuccessful bacterial transformations. It involves the use of a vector containing a gene for beta-galactosidase, which produces blue colonies when the desired gene is present. This method is simple and efficient, but may have limitations when the transformed gene does not produce beta-galactosidase.
  • #1
MrGenetic
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Books say 3 possibilities for this situation as insert within Lacz, insert outside Lacz, no inserted vectors, then these goes the transformation with Ecoli. But we can't know transformation yield %100. Thus, if we make blue white screening there is more possibilities.
Lacz-- inserted within white
Lacz-- inserted outside blue
Lacz-- no inserted blue
** No transformation white** ? ---> is this correct ? if it is correct, how we know the lacz inserted or not inserted clearly?
 
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  • #2
Usually the plasmid contains an antibiotic resistance marker, so that cells which do not get transformed will die and not produce any colony.
 
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Related to How do we know which transformation is occurring with blue/white screening?

1. How does blue/white screening work?

Blue/white screening is a technique used to distinguish between bacterial cells that have taken up a desired gene (transformants) and those that have not. It involves the use of a vector containing a gene for beta-galactosidase, an enzyme that converts a colorless substrate into a blue compound. Only bacteria with the desired gene will produce functional beta-galactosidase, resulting in blue colonies on a plate.

2. How do we know if the transformation was successful?

A successful transformation using blue/white screening will result in the formation of blue colonies on the plate. This indicates that the bacteria have taken up the desired gene and are producing functional beta-galactosidase.

3. Can blue/white screening be used to detect mutations in the transformed gene?

No, blue/white screening is only used to identify successful transformations. It cannot be used to detect mutations in the transformed gene. Additional methods such as DNA sequencing would be needed for this purpose.

4. What are the advantages of using blue/white screening?

Blue/white screening is a simple and efficient method for identifying successful transformations. It allows for the rapid screening of a large number of bacterial colonies and does not require specialized equipment or reagents.

5. Are there any limitations to using blue/white screening?

While blue/white screening is a useful technique, it is not always suitable for all types of gene cloning. For example, if the desired gene does not produce beta-galactosidase or if the bacteria naturally produce the enzyme, the results may be difficult to interpret. In these cases, alternative methods such as antibiotic resistance or fluorescent markers may be used for screening.

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