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Team:Grinnell/Notebook/Gels/DspB
From 2011.igem.org
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| - | Figure 9: Optimized <i>dspB + <i> rsaA</i> with P<sub>xyl</sub> and P<sub>rsaA</sub>. Lane 1: ladder; lanes 2-5: P<sub>rsaA</sub> + optimized <i>dspB</i> + <i>rsaA</i> colonies 1-4; lanes 6-9: P<sub>xyl</sub> + optimized <i>dspB</i> + <i>rsaA</i> colonis 1-4; lane 10: standard optimized <I>dspB</I> + <i>rsaA</i>. P<sub>rsaA</sub> + optimized <i>dspB</i> + <i>rsaA</i> colony 3 shows the expected size. | + | Figure 9: Optimized <i>dspB</i> + <i>rsaA</i> with P<sub>xyl</sub> and P<sub>rsaA</sub>. Lane 1: ladder; lanes 2-5: P<sub>rsaA</sub> + optimized <i>dspB</i> + <i>rsaA</i> colonies 1-4; lanes 6-9: P<sub>xyl</sub> + optimized <i>dspB</i> + <i>rsaA</i> colonis 1-4; lane 10: standard optimized <I>dspB</I> + <i>rsaA</i>. P<sub>rsaA</sub> + optimized <i>dspB</i> + <i>rsaA</i> colony 3 shows the expected size. |
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| - | Lane 1: ladder; lanes 2-5: | + | Figure 14: Lane 1: ladder; lanes 2-5: BBa_K081005, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lanes 6-9: P<sub>rsaA</sub>, optimized <I>dspB</i>, and <i>rsaA</i> in pMR10; lane 10: standard DNA fragment of optimized <i>dspB</i> with <i>rsaA</i>. çolony 2 carries P<sub>rsaA</sub> + <I>dspB</i> + <i>rsaA</i>. |
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| - | Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-6: PCR products; lane 7: standard fragment of <i>dspB</i> with <i>rsaA</i>. | + | Figure 15: Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-6: PCR products; lane 7: standard fragment of <i>dspB</i> with <i>rsaA</i>. |
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| - | Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-9: transformant PCR products; lane 10: digested miniprep of what should have been WT <i>dspB</i>, but showed up as the wrong fragment size. | + | Figure 16: Gel of PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i>. Lane 1: ladder; lanes 2-9: transformant PCR products; lane 10: digested miniprep of what should have been WT <i>dspB</i>, but showed up as the wrong fragment size. |
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| - | PCR of transformants that should carry BBa_K081005, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-7: PCR products. | + | Figure 17: PCR of transformants that should carry BBa_K081005, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-7: PCR products. |
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| - | PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-8: PCR products. | + | Figure 18: PCR of transformants that should carry P<sub>xyl</sub>, optimized <i>dspB</i>, and <i>rsaA</i> in pMR10. Lane 1: ladder; lanes 2-8: PCR products. |
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| - | Lane 1: ladder; lanes 2-9: colony PCR of transformants that should contain BBa_K081005, optimized <i>dspB</i>, and <i>rsaA</i>. | + | Figure 19: Lane 1: ladder; lanes 2-9: colony PCR of transformants that should contain BBa_K081005, optimized <i>dspB</i>, and <i>rsaA</i>. |
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Latest revision as of 14:10, 5 August 2011
Gels for DspB
2011.07.03-2011.07.09
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| Figure 1: Gel proving the successful transformation of WT dspB from UBC into E. coli Top10. Lane 1: ladder; lanes 2-4: digests (EcoRI and PstI) of miniprep DNA from overnights of transformant colonies 1-3. There is a clear band at the appropriate size for dspB in all of the experimental lanes. | Figure 2: Gel of transformants that should contain both WT dspB and a promoter insert. Lane 1: ladder; lanes 2-4: WT dspB with PrsaA colonies 1-4; lanes 5-7: WT dspB with Pxyl colonies 1-4; lane 8: standard dspB with no promoter. | Figure 3: Follow up to Figure 2. Gel showing WT dspB with various promoter inserts as contrasted to a standard dspB with no promoter. Lane 1: WT dspB + PrsaA colony 3; lane 2: with Pxyl colony 3; lanes 3,4: with BBa_K081005 colonies 3 and 4; lane 5: standard WT dspB; lane 6: ladder. | Figure 4: Test of more WT dspB with Pxyl colonies. Lane 1: ladder; lanes 2-5: WT dspB with Pxyl colonies 5-8; lane 6: standard dspB with no promoter. |
2011.07.10-2011.07.16
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| Figure 5: Optimized dspB transformed into pSB1C3 colonies A-E. Lane 1: ladder; lanes 2-5: optimized dspB colonies A-E; lanes 6-9: optimized esp. Colony D shows the expected size. | Figure 6: More colonies of WT dspB with Pxyl promoter. Lanes 1-4: colonies 9, 10, 12, and 15; lane 5: dspB WT standard from PCR. | Figure 7: The rest of the PCR product of colony 10 from Figure 6 in preparation for gel extraction of WT dspB behind Pxyl. Lane 1: ladder; lanes 2,3: WT dspB with Pxyl. | Figure 8: Gel of colony PCR of transformants carrying optimized dspB and rsaA C-terminal. Lane 1: ladder; lanes 2-5: transformation colonies 1-4; lane 6: dspB standard from PCR of WT dspB. |
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| Figure 9: Optimized dspB + rsaA with Pxyl and PrsaA. Lane 1: ladder; lanes 2-5: PrsaA + optimized dspB + rsaA colonies 1-4; lanes 6-9: Pxyl + optimized dspB + rsaA colonis 1-4; lane 10: standard optimized dspB + rsaA. PrsaA + optimized dspB + rsaA colony 3 shows the expected size. | Figure 10: Colonies 5-8 of Pxyl + dspB + rsaA. Lane1: ladder; lanes 2-5: carry Pxyl + optimized dspB + rsaA; lane 6: standard optimized dspB with rsaA from PCR. None of these colonies were successful. | Figure 11: Lane 1: standard optimized dspB with rsaA from PCR; lanes 2,3: transformants that should carry WT dspB behind Pxyl; lane 4: ladder. No successful transformations. | Figure 12: BBa_K081005 + dspB + rsaA colonies 1-4. Lane 1: ladder; lanes 2-5: BBa_K081005 + optimized dspB + rsaA; lane 6: standard optimized dspB with rsaA fragment. Colony 2 is successful. |
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| Figure 13: Pxyl + dspB + rsaA colonies 9-14. Lane 1: ladder; lanes 2-7: Pxyl + dspB + rsaA; lane 8: standard fragment of optimized dspB with rsaA. |
2011.07.17-2011.07.23
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| Figure 14: Lane 1: ladder; lanes 2-5: BBa_K081005, optimized dspB, and rsaA in pMR10; lanes 6-9: PrsaA, optimized dspB, and rsaA in pMR10; lane 10: standard DNA fragment of optimized dspB with rsaA. çolony 2 carries PrsaA + dspB + rsaA. | Figure 15: Gel of PCR of transformants that should carry Pxyl, optimized dspB, and rsaA. Lane 1: ladder; lanes 2-6: PCR products; lane 7: standard fragment of dspB with rsaA. | Figure 16: Gel of PCR of transformants that should carry Pxyl, optimized dspB, and rsaA. Lane 1: ladder; lanes 2-9: transformant PCR products; lane 10: digested miniprep of what should have been WT dspB, but showed up as the wrong fragment size. | Figure 17: PCR of transformants that should carry BBa_K081005, optimized dspB, and rsaA in pMR10. Lane 1: ladder; lanes 2-7: PCR products. |
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| Figure 18: PCR of transformants that should carry Pxyl, optimized dspB, and rsaA in pMR10. Lane 1: ladder; lanes 2-8: PCR products. |
2011.07.24-2011.07.31
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| Figure 19: Lane 1: ladder; lanes 2-9: colony PCR of transformants that should contain BBa_K081005, optimized dspB, and rsaA. |
