BIOLOGY NOW(LL)(FDS)
2nd Edition
ISBN: 9780393683790
Author: HOUTMAN
Publisher: NORTON
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Chapter 6, Problem 4TQ
Summary Introduction
The cell cycle is an important mechanism used to increase the cell number while maintaining genetic stability of cells. Cell cycle plays a crucial role in growth as well as regeneration. Interphase and cell division are two important stages occur in the cell cycle. Mitosis (maintains the total number of chromosomes) and meiosis (reduces half the chromosome number) are two kinds of cell divisions occur in eukaryotes. Prokaryotes use the mechanism of binary fission (a sexual reproduction) for cell division.
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Overview of Transformation Protocol
-Prepare competent bacteria for transformation:
Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.
Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.
-Transformation procedure:
Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.
Add CaCl2 to both tubes.
Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.
Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.
Add recovery broth to both tubes.
Incubate both tubes in a 37 C water bath for 5 minutes.
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