You are given a 10 X protein gel running buffer (Tris/Glycine/SDS). Make 800 ml

Question

You are given a 10 X protein gel running buffer (Tris/Glycine/SDS). Make 800 ml for electrophoresis of your samples on a SDS-PAGE gel. 10. You have 10 uL off protein samples to analyze for induced bacteria. You need to boil samples and perform SDS-PAGE You have 3X Laemmli buffer. Show how you would process samples for the loading. BONUS: IPTG is an inducer. One can also use _______. The advantage of using IPTG is that ___________ By what general methodology were the bacterial cells lysed after induction ?_________ What other method could have been used? __________

Explanation / Answer

(9)

Stock: 10x

Required: 1x

Volume : 800 mL

C1V1= C2V2

(stock)   (required)

V1=C2V2/C1

V1 = ( 1x X 800 mL )/ 10x

= 80 mL

Take 80 mL + 720 mL of H2O to get 1x running buffer

(10)

10uL samples

3x Laemmli buffer

Final volume 10uL

We need final 1x laemmli

Aliquot vol =(aliquot volume + diluent volume)/dilution factor

= 10uL/3

= 3.33

To load=Take 3.3uL of 3x Laemmli buffer + 6.7uL of protein sample

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