Brdu incorporation principle

the principle of brdu incorporation is that after BrdU is added to the culture medium, it can be used as the raw material for cell DNA replication. After two cell cycles, the DNA with two single strands of BrdU in the cell will account for l/2, which reflects that it should be a single monomer lightly stained on the chromosome.

after three cycles, about half of the chromosomes are light stained with two monomers, and the other half are dark and light. If the cell only goes through one cycle, both monomers are deeply stained. The value of cell cycle can be calculated by counting the proportion of each phase in the mitotic phase.

cell cycle refers to the time that a generation of cells has passed. It is a cycle from the end of one cell division to the end of the next. Cell cycle reflects the rate of cell proliferation. Time-lapse photography can be used to measure the cycle of a single cell, but it cannot represent the cycle of a cell population, so other methods are often used to measure the cycle of a population. Brdu incorporation principle is a method to measure cell cycle.

operation steps of BrdU:

1. When the cells grow to exponential stage, add BrdU to the culture medium to make the final concentration 1μg/ml.

colchicine was added in 2 hours and 44 hours to make each ml contain .1 μ g.

After 3 or 48 hours, digest the cells into the centrifuge tube, and pay attention to the floating cells in the culture supernatant.

4. Routine chromosome production (see Part III: Chromosome Technology).

5. Put the chromosome slide on the cover of a 56℃ water bath pot, spread 2×SSC solution, and irradiate it with ultraviolet light at a distance of 6cm from the ultraviolet lamp for 3 minutes.

6. discard the 2×SSC solution and rinse with running water.

7. Dye with Giemsa solution for 1 minutes, rinse with running water and dry.

8. Microscopically examine 1 mitotic phases and record the mitotic index of the first, second, third and fourth cell stages.

what are the above references? Baidu encyclopedia -BrdU