Canadian BioSample Repository

SITE: Laboratory (CL) CATEGORY: Reagent Preparation (02) PREPARED BY: Jonathan Hooton APPROVED BY: SIGNED (BLUE INK): DATE SIGNED:

CBSR

PBS (Phosphate Buffered Saline) pH 7.3

INTRODUCTION This isotonic solution is used in the preparation of whole human blood cells. It is prepared as a 10x solution, diluted and autoclaved. SOURCE Irene AMENDMENTS FROM PREVIOUS DOCUMENT None DOCUMENTS Work sheet: Title: Document #: PBS pH 7.3 Worksheet CL02AM00--

MATERIALS Reagent bottle: Supplier: Catalogue #: Capacity: Location: Location: Location: Location:

500 ml balance drawer glassware cupboard glassware cupboard glassware cupboard

Magnetic stir bar: 500 ml beaker: 5 Litre beaker: 500 ml cylinder: REAGENTS NaCl:

Product: Manufacturer: Supplier: Catalogue #: Weight: MW: Location:

Sodium Chloride EM Science Biochemistry Stores, Rm 471 MSB 5X0420-3 2.5 kg 58.44 Reagent shelf

Version Date: Print Date: Page:

4/28/2003 1/5/2006 1 of 4

Document control #: Controlled copy #: Document type:

1. Master SOP

This document is a confidential communication of the Canadian BioSample Repository Receipt of this document constitutes an agreement by the recipient that no unpublished information contained herein shall be published or disclosed without written approval, except that this document may be disclosed to the appropriate REB under the condition that they keep it confidential.

Canadian BioSample Repository

SITE: Laboratory (CL) CATEGORY: Reagent Preparation (02) PREPARED BY: Jonathan Hooton KCl: Product: Manufacturer: Supplier: Catalogue #: Weight: MW: Location: Product: Manufacturer: Supplier: Catalogue #: Weight: MW: Location: Product: Manufacturer: Supplier: Catalogue #: Weight: MW: Location: Supplier: APPROVED BY: SIGNED (BLUE INK): DATE SIGNED: Potassium Chloride EM Science Biochemistry Stores, Rm 471 MSB PX1405-5 2.5 kg 74.56 Reagent shelf Disodium hydrogen phosphate Anachemia Biochemistry Stores, Rm 471 MSB 84486-380 2 kg 141.96 Reagent shelf Sodium dihydrogen phosphate EM Science Biochemistry Stores, Rm 471 MSB PX1565-5 2.5 kg 136.09 Reagent shelf Bleackley lab

CBSR

Na2HP04:

KH2P04:

Double distilled water:

INSTRUMENTS 1) Mettler College weighing balance 2) Fisher pH meter PREPARATION 01) Obtain next sequential worksheet 02) Enter your name and the date on the work sheet. 03) Enter lot numbers of NaCl, KCl, KH2P04 and KH2P04 on the work sheet. 04) Weigh out 41.62 g NaCl (1.42 mM final concentration) in the beaker on weighing balance and enter weight in box on work sheet. 05) Weigh out 1.04 g KCl in a small weighing boat and enter weight in box on worksheet 06) Weigh out 5.21 g Na2HP04 into a medium size weighing boat and enter weight in box on worksheet.
Version Date: Print Date: Page: 4/28/2003 1/5/2006 2 of 4 Document control #: Controlled copy #: Document type:

1. Master SOP

This document is a confidential communication of the Canadian BioSample Repository Receipt of this document constitutes an agreement by the recipient that no unpublished information contained herein shall be published or disclosed without written approval, except that this document may be disclosed to the appropriate REB under the condition that they keep it confidential.

Canadian BioSample Repository

SITE: Laboratory (CL) CATEGORY: Reagent Preparation (02) PREPARED BY: Jonathan Hooton APPROVED BY: SIGNED (BLUE INK): DATE SIGNED:

CBSR

07) Weigh out 1.51 g KH2PO4 into a small weighing boat and note weight in box on worksheet. 08) Add 350 ml distilled water to the beaker. 09) Pour KCl, KH2P04 and KH2P04 into the beaker and wash any remnants into beaker with distilled water from a squirt bottle. 10) Place large stir bar in beaker, place beaker on magnetic stirrer and start stirrer. 11) When all the solid material has dissolved, adjust the pH to 7.3 with 1 N NaOH solution. 12) Pour solution into 500 ml graduated cylinder. Add distilled water to 500 ml mark. 13) Cover top of cylinder with parafilm and mix by inverting the cylinder. 14) Pour solution into 5 litre beaker and add 4.5 litre H20 using 2 L graduated cylinder, washing the solution from the 500 ml cylinder into the 5 L beaker. 15) Place large stir bar in the beaker and mix the solution on the magnetic mixer. 16) Pour solution into ten 500 ml reagent bottles screw on caps lightly. 17) Write your initials and reagent id on unused autoclave and attach to bottle. 18) Autoclave bottles in tray with water for 20 minutes, at 1220 C. 19) After removing from bottles from autoclave screw on caps tightly and leave to cool. 20) Perform validation checks. 21) Enter results of validation checks on work sheet. 22) Indicate whether batch accepted or rejected. 23) Sign work sheet and obtain supervisors signature. 24) Place work sheet in CL02AM binder. 25) Label each bottle with: BPP-CL

PBS
Phosphate Buffered Saline pH 7.3 STERILE Store at room temperature Prep Date Batch e # Expiry Date

22) Enter present date under Prep Date 23) Enter worksheet document control number under Batch #. 24) Calculate expiry date from shelf life given below. VALIDATION 1) pH. a) Using sterile technique, take a 10 ml sample of PBS. b) Measure pH.
Version Date: Print Date: Page: 4/28/2003 1/5/2006 3 of 4 Document control #: Controlled copy #: Document type:

1. Master SOP

This document is a confidential communication of the Canadian BioSample Repository Receipt of this document constitutes an agreement by the recipient that no unpublished information contained herein shall be published or disclosed without written approval, except that this document may be disclosed to the appropriate REB under the condition that they keep it confidential.

Canadian BioSample Repository

SITE: Laboratory (CL) CATEGORY: Reagent Preparation (02) PREPARED BY: Jonathan Hooton c) Enter result on work sheet. Reject whole batch if pH < 7.2 or > 7.4 STORAGE Store at room temperature on reagent shelf. Store at 40 C in fridge when bottle opened. SHELF LIFE 24 months at room temperature if unopened. 4 weeks at 40 C when opened. APPROVED BY: SIGNED (BLUE INK): DATE SIGNED:

CBSR

Version Date: Print Date: Page:

4/28/2003 1/5/2006 4 of 4

Document control #: Controlled copy #: Document type:

1. Master SOP

This document is a confidential communication of the Canadian BioSample Repository Receipt of this document constitutes an agreement by the recipient that no unpublished information contained herein shall be published or disclosed without written approval, except that this document may be disclosed to the appropriate REB under the condition that they keep it confidential.