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Myocyte digestion buffer
 

Reagent name: Myocyte digestion buffer

Reagent name abbreviation: MC digestion buffer

Protocol ID: PS00000447

Version: 01

Volume: 50 ml

Components:

Reagent

Source

Catalog or Protocol No.

F.W. or Stock Concentration

Quantity

Final Concentration

Perfusion buffer

 

None

 

PS00000451

 

1X

 

50 mg

1X

 

Liberase

blendzyme 1

 

Roche

 

1988417

 

NA

 

12.5 mg

 

0.25 mg/ml

 

Trypsin

 

GIBCO/

Invitrogen

 

15090046

 

10X

 

0.278 ml

 

0.14 mg/ml

 

Calcium chloride

(CaCl2), 100 mM

 

None

 

PS00000441

 

100 mM

 

6.25 µl

 

12.5 µM

 

Preparation:

1. Weigh liberase blendzyme 1 and add to 30 ml of perfusion buffer in a 100-ml beaker.

2. Measure and add trypsin and 100 mM CaCl2 to the beaker.

3. Stir until all components are thoroughly dissolved.

4. Adjust final volume to 50 ml with perfusion buffer in a graduated cylinder.

5. Cover with Parafilm and mix by inversion.

6. Prewash a 0.2 µM filter attached to a 60-ml syringe with 10 ml of purified water.

7. Sterilize the solution by filtering through the prewashed filter.

Storage:

Temperature: NA

Location: __________________

Aliquot size: NA

Special instructions: Do not store solution. Prepare fresh as needed.

Author:

Date:    

Approved:

Comments:

 

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