Agarose Electrophoresis Gel Preparation and Practice

Only available on StudyMode
  • Topic: Agarose gel electrophoresis, Gel electrophoresis, Tris
  • Pages : 3 (561 words )
  • Download(s) : 490
  • Published : February 10, 2012
Open Document
Text Preview
AGAROSE ELECTROPHORESIS GEL PREPARATION AND PRACTICE SAMPLE LOADING STANDARD OPRERATING PROCEDURE

ID NUMBER 1PREPARDED BY CHRIS FOLEY
REVISION NUMBER 1PAGE 1 OF 3
DATE OF REVISION 2-1-12

1. STATEMENT OF PURPOSE
Agarose Electrophoresis gel preparation and practice.

2. SCOPE
Prepare a SOP for the preparation of 1% agarose gel using TBE buffer and practice electrophoresis gel loading procedure.

3. STATEMENT OF RESPONSIBILITY
This procedure has been carried out by Chris Foley

4. MATERIALS REQUIRED
Calculatorweighting boats
Scale1000ml beaker
Spatula500ml graduated cylinder
Magnetic stirrerstir bar
Hot plateparafilm
Electrophoresis chambergel bed with well former
Micropipette tipssample loading solution
Distilled waterlabeling tape
Gogglesgloves
250 ml glass flaskthermometer

5. REAGENTS AND CHEMICALS
A. Tris BaseManufacturerFisher Scientific
Lot Number976707A
Formula Weight121.14 grams

B. Boric AcidManufacturerFisher Scientific
Lot Number99715
Molecular Weight61.83grams

SOP AGROSE GEL ELECTROPHORESIS PREP & PRACTICEPAGE 2 OF 22-1-12 C. EDTAManufacturerFisher Scientific
Lot Number 073458
Formula Weight372.24grams

D. AgaroseEdvotek
Lot LF45100003
6. CALCULATIONS
Calculations for preparing the grams needed to make 500ml of Trisborate buffer using concentrations of reagents listed below. Calculate grams of solute multiply ml time’s mole time’s molecular weight. A. 0.89mM Tris Base

0.00089m x .500l x 121.14g/m = 0.053 grams needed

B. 0.89mM Boric Acid
0.00089m x .500l x 61.83g/m = 0.027 grams needed

C. 2.5mM Na2EDTA
.0025m x .500l x 372.24g/m = 0.465 grams needed

7. PROCEDURE
A. Buffer Preparation
1. Place 1000ml beaker onto stir plate containing 60ml of distilled water and magnetic stirrer, turn on stir plate 2. Add above calculated...
tracking img