Question

You determined in the previous assignment the exact volume of 50X TAE that you will need. I suggest that you make 40 ml of 50X TAE so that you have some extra (just in case you need it). The recipe for one liter of 50X TAE is as follows: ¥ 242g Tris base ¥ 57.1 ml glacial acetic acid ¥ 100 ml 0.5 M EDTA pH 8.0 9. Provide a recipe for 40 ml of 50X TAE. 10. When you make your agarose gel, you will need to make 50 ml of 1% (w/v) agarose in 1X TAE. How much agarose will you require? [you will make this later, not on Solution day]

Answer 1

Hi Answer:

[The below given calculations are the simplest calculations to understand how the volume reduces with change in concentration so try to understand step by step]

As the recipe of 1 litre, 50X TAE is given in question i.e.

50X TAE (1 L)

242 g Tris base, 57.1 mL Glacial acetic acid, 100 mL 0.5 M EDTA pH 8.0

DD water 842.9 mL (this volume comes from Glacial acetic acid & EDTA volume – 1 L (total volume we need)).

So all components needed for 40 mL volume are:

Tris base needed for 40mL = 242/50 =4.84/1000=0.00484 X 40= 0.1936 g

Glacial acetic acid needed for 40 mL= 57.1/50= 1.142/1000 =0.001142 X 40 =0.04568 mL

EDTA needed for 40 mL= 100/50 =2/1000 =0.002X40 =0.08 mL

AND 39.8744 mL of water to make the 40 mL (1X TAE) solution

[In above calculation we divide in first step with 50 to make 1X, then divide with 1000 to find the amount/volume in 1 mL and at last multiply by 40 to make final volume 40 mL].

Next to make 50 mL of 1X TAE

Simply we make 50 mL of TAE form above calculations.

Tris base needed for 50mL = 0.00484 X50= 0.242 g

Glacial acetic acid needed for 40 mL=0.001142 X 50 =0.0571mL

EDTA needed for 40 mL= 0.002X50 =0.1 mL

AND 49.8429 mL of water to make the 50 mL (1X TAE) solution.

OR you can preapre 1X TAE (50mL) from above given 50X TAE solution by taking 1 mL (50X TAE) and mix it with 49 mL of water. The resultant TAE solution has 1X concentration

Now add 1% agarose gel means 1 g/100 mL and we need 50 mL simple add 0.5 gram of agarose in 50 mL of (1X) TAE. This concentration of gel is 1% in 50 mL of 1X TAE