OVERVIEW
A 0.1 mg/ml Poly-d-lysine solution is used to coat coverslips. Coated coverslips allow cells to adhere and grow. A 0.1 mg/ml Poly-d-lysine solution can be prepared either from the solid poly-d-lysine or by diluting the 1 mg/ml stock solution.
Here we describe a procedure to prepare a 0.1 mg/ml stock solution of Poly-d-lysine by diluting the 1 mg/ml stock solution.
REQUIREMENTS
Reagents and solutions
♦ 1 mg/ml Poly-D-Lysine hydrobromide solution
♦ MilliQ water (autoclaved sterile)
Equipment and disposables
♦ Pipette and pipette boy
♦ Micropipette and tips
♦ Falcon® screw cap graduated Conical Tubes, 15 mL (e.g., SARSTEDT, 62.554.502)
♦ Laminar flow hood
COMPOSITION
0.1 mg/ml Poly-d-lysine hydrobromide solution in water
OBJECTIVE
Preparation of 0.1 mg/ml Poly-D-Lysine solution from 1 mg/ml stock solution
Use appropriate personal protective equipment (lab coat, gloves, goggles, etc) for your safety and follow your institute’s guidelines.
Prior to start:
Make the laminar floor hood ready for work (switch on UV for 20 min, and then start the airflow)
Step 1: Clean all the reagent bottles and disposables (1 mg/ml Poly-d-lysine hydrobromide stock solution bottle, autoclaved MilliQ water bottle, 15 mL Falcon® screw cap graduated Conical Tubes) with 70% ethanol and bring them inside the laminar flow hood.
Note:
To clean the outer surface of the reagent’s bottles, 15 ml tube, pipette boy and micropipette, and tip boxes, spray 70% ethanol on them and wipe their surfaces with tissue paper. Take extra care while spraying and cleaning the lid portion of the bottle. This will help you keep the poly-d-lysine solution sterile.
Step 2: Transfer 1 ml poly-d-lysine stock solution from the stock bottle to a 15 ml Falcon® screw cap graduated Conical Tube. Add 9 ml MilliQ water. Screw the cap tightly and mix the solution by inverting the tube a few times.
STORAGE
Store the 0.1 mg/ml poly-d-lysine solution at 4°C.
