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SL-5’UTR control – 30’. SL-5’UTR control – 0’. SL-AUG control – 30’. SL-AUG control – 0’. 5’UTR control – 30’. 5’UTR control – 0’. AUG control – 30’. AUG control – 0’. 28S rRNA. 16S rRNA. S1: Homogeneity and stability of the mRNA translated: - PowerPoint PPT Presentation
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28S rRNA
16S rRNA
5’U
TR
co
ntr
ol –
0’
SL
-5’U
TR
co
ntr
ol –
0’
SL
-AU
G c
on
tro
l – 3
0’
AU
G c
on
tro
l – 3
0’
SL
-AU
G c
on
tro
l – 0
’
AU
G c
on
tro
l – 0
’
SL
-5’U
TR
co
ntr
ol –
30’
5’U
TR
co
ntr
ol –
30’
S1: Homogeneity and stability of the mRNA translated:A - Homogeneity and stability of the mRNA constructs in the RRL. 0.05 µM of 32P labeled 5’UTR Control, SL-5’UTR Control, AUG Control, SL-AUG control were incubated for 0 or 30 minutes in 15 µL RRL under
translational conditions. The reaction was stopped on ice in the presence of 1% SDS, phenol extracted, ethanol precipitated, and the RNAs analysed on a 1% denaturing agarose gel. The 18S and 28S rRNA were visualized by SyBr staining before autoradiography and are indicated on the left hand side of the figure.
HIV
-2-5
’UT
R –
0’
SL
-HIV
-2-5
’UT
R –
0’
SL
-HIV
-2-A
UG
1 –
0’
HIV
-2-A
UG
1 –
0’
28S rRNA
16S rRNA
HIV
-2-A
UG
1 –3
0’
SL
-HIV
-2-A
UG
1 –
30’
SL
-HIV
-2-5
’UT
R –
30’
HIV
-2-5
’UT
R –
30’
B - Homogeneity and stability of the mRNA constructs in the RRL. 0.05 µM of 32P labeled HIV-2-5’UTR, SL-HIV-2-5’UTR, HIV-2-AUG1, SL-HIV-2-AUG1 were incubated for 0 or 30 minutes in 15 µL RRL under translational conditions. The reaction was
stopped on ice in the presence of 1% SDS, phenol extracted, ethanol precipitated, and the RNAs analysed on a 1% denaturing agarose gel. The 18S and 28S rRNA were visualized by SyBr staining before autoradiography and are indicated on the left hand side of the figure.
