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

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SUMMARY

The discussion focuses on the complexities of blue/white screening in E. coli transformation, specifically regarding the Lacz gene's insertion. Three scenarios are identified: Lacz inserted within the white colony, Lacz inserted outside the blue colony, and no insertion resulting in blue colonies. The uncertainty of transformation yield is emphasized, as it is impossible to determine with 100% certainty whether Lacz has been inserted. The presence of an antibiotic resistance marker in the plasmid is crucial, as it ensures that only transformed cells survive and form colonies.

PREREQUISITES
  • Understanding of E. coli transformation techniques
  • Knowledge of blue/white screening methodology
  • Familiarity with the Lacz gene and its role in plasmid constructs
  • Basic concepts of antibiotic resistance markers in molecular biology
NEXT STEPS
  • Research the mechanisms of blue/white screening in E. coli
  • Learn about the role of antibiotic resistance markers in plasmid selection
  • Explore methods to confirm Lacz insertion using PCR
  • Investigate alternative screening techniques for plasmid transformation
USEFUL FOR

Molecular biologists, genetic engineers, and researchers involved in plasmid construction and E. coli transformation will benefit from this discussion.

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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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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