karyotyping- Step and procedure

How to do Karyotyping?

Sample collection, sample processing, cell culture, incubation, cell harvesting, slide preparation, microscopy and preparing a karyogram are common steps in karyotyping.

Steps in Karyotyping:

Collect the Blood sample in the Heparin tube and Mix well at room temperature.

Transport the sample Under the appropriate condition, usually, at 4°C, immediately withing 24 hours of collection.

Pre-process of the sample before karyotyping to separate leukocytes.

Perform cell culture using the ready to use RPMI-1640 media under strict aseptic conditions.

Incubate sample in a CO2 incubator for 72 hours at 37°C temperature.

Perform cell harvesting using repeated centrifugation until clean pallets observed.

Prepare a slide from the culture, drop some liquid above two feet hight, and stain with Giemsa stain.

Perform GTG banding using the Giemsa-Trypsin-Giemsa protocol to visualize chromosomes.

Observe the slide under the microscope in 10X, 45X and 100X to investigate chromosomal abnormalities.

Arrange chromosomes in an orderly manner to prepare a karyogram.