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Post Type: SDS_PAGE
A 12% SDS-PAGE gel was polymerised according to Sambrook. Samples were loaded in gel loading buffer and the gel run at 80 V for approximately 1 hour.
The gel was first imaged under UV light (short wave band emission filter) and then stained with coomassie.
Lane | Sample | ul |
1 | ... | |
2 | ... | |
3 | Protein MW Marker | 5 |
4 | ... | |
5 | GG-TerB-BFP conjugate | 5 |
6 | ... | |
7 | ds-GG-TerB- BFP conjugate | 5 |
8 | ... | |
9 | Product of ligation of GG-Fluor to Tus | 5 |
10 | ... | |
11 | ... | |
12 | ... | |
13 | ... | |
14 | ... | |
15 | ... | |
16 | ... |
A 12% SDS-PAGE gel was polymerised according to Sambrook. Samples were loaded in gel loading buffer and the gel run at 80 V for approximately 1 hour.
The gel was first imaged under UV light (short wave band emission filter) and then stained with coomassie.
Attached Files
Fluorescent imaging of gel
Fluorescent image of gel
Coomassie stained gel
Coomassie stained gel
The fluorescence image shows that the Tus protein has been successfully labelled with fluorescein. The small band higher up is the protein, whereas the larger fluroescent band is the unligated label.
The coomassie gel shows that we have successfully ligated both double and single stranded DNA to the BFP protein. The lower band in lanes 5 and 7 (doublet) is unmodified BFP (runs near the 30 kDa marker) whereas the upper band is the presumed conjugate. Single and double stranded conjugate are expected to run at the same position as the second strand would be stripped off the double stranded conjugate.
The coomassie gel shows that we have successfully ligated both double and single stranded DNA to the BFP protein. The lower band in lanes 5 and 7 (doublet) is unmodified BFP (runs near the 30 kDa marker) whereas the upper band is the presumed conjugate. Single and double stranded conjugate are expected to run at the same position as the second strand would be stripped off the double stranded conjugate.