CELL FREE SYSTEMS	> Wheat Germ Cell-Free Translation	> Protocol for wheat germ cell-free translation
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		> Preparation of a wheat germ extract

 

Sample WG Protocol

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Translating various molecules with different amounts of microsomes.

WG master mix For 10 uL	For 330 uL experiment
CB 10X 1 uL	33 uL
E mix 2	66
WG 2	66
tRNA 0.1	3.3
CK 0.1	3.3
RNasin 0.1	3.3
Water 3.2	106

Translations:

Tube Transcript Volume MM KRM RM H2O	Total
1 SgGSTP -mb 4 15 - - 1	20
2 SgGSTP +mb 10 37.5 - 2.5 -	50
3 SgGSP -mb 4 15 - - 1	20
4 SgGSP +mb 10 37.5 - 2.5 -	50
5 SG1 -mb 2 7.5 - - 0.5	10
6 SG1 +mb 4 15 - 1 -	20
7 125C -mb 2 7.5 - - 0.5	10
8 125C +mb 4 15 - 1 -	20
9 pSPgG8 -mb 2 7.5 - - 0.5	10
10 pSPgG +mb1 4 15 - 0.5 0.5	20
11 pSPgG +mb2 4 15 - 1 -	20
12 pSPSTgG -mb 4 15 - 0.5 0.5	10
13 pSPSTgG +mb1 4 15 - 0.5 0.5	20
14 pSPSTgG +mb2 4 15 - 1 -	20
15 SG1 + KRM 2 7.5 0.5 - -	10