MICROBIOLOGY FOR STUDENTS (M4S)

Chromosomal DNA Extraction Procedure Chromosomal DNA Extraction Procedure for Gram-Negative Bacteria Steps Description 1. Make about 20 ml of LB broth in a 50 ml test-tube for each isolate. 2. Inoculate appropriate loopful of desired microorganism in the LB broth and incubate for 24-48 hrs, depending upon the turbidity appearance. 3. Take 1.5ml of fresh culture (LB) broth in Eppendorf or microfuge tube with the help of pipette. 4. Centrifuge it at 5000 rpm for 10 min. 5. Repeat the same process for double harvest of cell mass by discarding the supernatant. 6. Add 200 microliter of Lysis buffer I and 10 microliter of lysozyme (0.5mg/ml). 7. Vortex the mixture. 8. Incubate the Eppendorf tube containing mixture at 37°C for 30 minutes. 9. Add 1/10th volume of 10% SDS (about 20 microliter). 10. Incubate it at...

The parC gene, also known as topoisomerase IV subunit A, is a bacterial gene that codes for a subunit of the topoisomerase IV enzyme. Topoisomerase IV is a type II topoisomerase enzyme that plays a crucial role in bacterial DNA replication, transcription, and recombination by resolving DNA topological problems, such as catenanes and knots, during chromosome segregation and cell division. Here's an overview of the parC gene and its functions: Gene Structure : The parC gene is typically found in the bacterial chromosome and is composed of a specific DNA sequence that encodes the ParC protein. Similar to other bacterial genes, the parC gene consists of coding regions (exons) that specify the amino acid sequence of the ParC protein, as well as non-coding regions (introns) that may contain regulatory elements involved in gene expression. Protein Function : The ParC protein is one of the subunits of topoisomerase IV, which functions as a DNA topoisomerase enzyme in bacteria. Topoisomeras...

The gyrA gene, also known as DNA gyrase subunit A, is a bacterial gene that codes for a subunit of the DNA gyrase enzyme. DNA gyrase is a type II topoisomerase enzyme that plays a crucial role in bacterial DNA replication, transcription, and recombination by introducing negative supercoils into DNA strands. Here's an overview of the gyrA gene and its functions: Gene Structure : The gyrA gene is typically found in the bacterial chromosome and is composed of a specific DNA sequence that encodes the GyrA protein. The gene consists of coding regions (exons) that specify the amino acid sequence of the GyrA protein, as well as non-coding regions (introns) that may contain regulatory elements involved in gene expression. Protein Function : The GyrA protein is one of the subunits of DNA gyrase, which functions as a DNA topoisomerase enzyme in bacteria. DNA gyrase plays a critical role in DNA replication by introducing negative supercoils ahead of the replication fork, thereby relieving the...