Pancreatin-Giemsa Banding

[EBJ]

Does anyone in the group have any experience and/or recommendations
for using pancreatin instead of trypsin for G-banding? I'm mostly
interested in peripheral blood and oncology specimen slides.

Thanks!

Eric

Eric B. Johnson
UW Cytogenetic Services
Madison, WI
http://slh.wisc.edu/cytogenetics/

[trypanblue]

Hi EBJ,
we have tried pancreatic G-Banding in the past and have found the
results similar to trypsin G-Banding. There is a method below for you
to try.

Preparing Pancreatin Solution for G-Banding:

Preset a waterbath at 37°C.
To make 1X Hanks solution, place 5 ml of Hanks solution (10X) and
40.5
ml of distilled water in a
coplin jar. Stir to mix. Heat this solution to 37°C by placing the
coplin jar in the waterbath
(about 20 minutes). Add 50 mg pancreatin to 2 ml distilled water and
mix. Add the pacreatin
solution to the 1X Hanks solution and mix. Then, using a Pasteur
pipet, carefully add 1 or more
drops of 7.5% sodium bicarbonate solution, mixing after each drop to
achieve orange-red color
(not pink). The pH of this solution is critical. Keep this coplin jar
in the waterbath at 37°C.

G-Banding Method using Pancreatin:

1. Place slides in a coplin jar with previously prepared pancreatin
solution (see above) in the
waterbath at 37°C. Incubate for 12 seconds.
2. Rinse the slide immediately in a tap water by dipping the slide in
and out of
water several times.
3. Shake and drain the excess water and place the slide in the Giemsa
stain solution.
Stain the chromosomes for 8–10 minutes.
4. Rinse the slide thoroughly in tap water by dipping it in and out
of
the
water several times.
5. Shake and drain the slide with a paper towel on the back side of
the slide.
6. After air-drying or heating the slide on a slide warmer (40–60°C)
for 1 minute, observe the slide
with a low power (10X) objective.
Ref:
C.C. Tseng
Department of Biological Sciences
Purdue University-Calumet
Hammond, Indiana 46323-2094