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Team:BostonU/Data
From 2014.igem.org
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<td scope="col">According to literature, the pTet-pBad construct has not previously functioned as suspected. This was possibly due to position-dependence interference [1]. Conversely, our flow cytometer data showed that pTet-pBad had a greater range of fluorescence than pBad-pTet. We will need to do further investigating to find out why this was the case. </td> | <td scope="col">According to literature, the pTet-pBad construct has not previously functioned as suspected. This was possibly due to position-dependence interference [1]. Conversely, our flow cytometer data showed that pTet-pBad had a greater range of fluorescence than pBad-pTet. We will need to do further investigating to find out why this was the case. </td> | ||
| - | <td scope = "col"><img src="https://static.igem.org/mediawiki/2014/3/39/PBad_pTet_RFP_all_three.png" width=" | + | <td scope = "col"><img src="https://static.igem.org/mediawiki/2014/3/39/PBad_pTet_RFP_all_three.png" height="500" width="500" style="float:right" style= "margin-left:10px"></td></tr></table> |
<br><br> | <br><br> | ||
<p><center><img src="https://static.igem.org/mediawiki/2014/3/34/Origins_graph.png" width="50%"></center></p><br><br> | <p><center><img src="https://static.igem.org/mediawiki/2014/3/34/Origins_graph.png" width="50%"></center></p><br><br> | ||
Revision as of 02:24, 16 October 2014
Primer Design for Tandem Promoters and Repressor Genes
| Device Name | Forward Primer | Sequence | Reverse Primer | Sequence |
|---|---|---|---|---|
| BetI_CD | BetI_For_C | ATGAAGACGTAATGGTGCCGAAACTGGGTATGCAGAGC | BetI_Rev_D | ACGAAGACCTACCTTTAATCGGTCGGCAGATGCTGGGT |
| PhlF_CD | PhlF_For_C | ATGAAGACGTAATGATGGCACGTACCCCGAGCCGTAGC | PhlF_Rev_D | ACGAAGACCTACCTTTAACGCTGTGTACCCGGACAAAC |
| BM3R1_CD | BM3R1_For_C | ATGAAGACGTAATGATGGAAAGCACCCCGACCAAACAG | BM3R1_Rev_D | ACGAAGACCTACCTTTAGCTCTGACGGCTCAGTGCTGC |
| LmrA_CD | LmrA_For_C | ATGAAGACGTAATGATGAGCTATGGTGATAGCCGTGAA | LmrA_Rev_D | ACGAAGACCTACCTTTAACGTTTCAGCAGATCCGGAAT |
| SrpR_CD | SrpR_For_C | ATGAAGACGTAATGATGGCACGTAAAACCGCAGCAGAA | SrpR_Rev_D | ACGAAGACCTACCTTTATTCGAAGGATTTCACCTGTTT |
| pTet_AK | pTet_For_A | ATGAAGACGTGGAGTCCCTATCAGTGATAGAGATTGAC | pTet_Rev_K | ACGAAGACCTGCATTTCGGTCAGTGCGTCCTGCTGATG |
| pTet_KB | pTet_For_K | ATGAAGACGTATGCTCCCTATCAGTGATAGAGATTGAC | pTet_Rev_B | ACGAAGACCTAGTATTCGGTCAGTGCGTCCTGCTGATG |
| pBad_AK | pBad_For_A | ATGAAGACGTGGAGAAGAAACCAATTGTCCATATTGCA | pBad_Rev_K | ACGAAGACCTGCATTATGGAGAAACAGTAGAGAGTTGC |
| pBad_KB | pBad_For_K | ATGAAGACGTATGCAAGAAACCAATTGTCCATATTGCA | pBad_Rev_B | ACGAAGACCTAGTATATGGAGAAACAGTAGAGAGTTGC |
| pSrpR_KB | pSrpR_For_K | ATGAAGACGTATGCTTCGTTACCAATTGACAGCTAGCT | pSrpR_Rev_B | ACGAAGACCTAGTAGTTTACAAACAAACAAGCATGTAT |
| pLmrA_FK | pLmrA_For_F | ATGAAGACGTCGCTTTCGTTACCAATTGACAACTGGTG | pLmrA_Rev_K | ACGAAGACCTGCATAAATATAGTGACTGGTCTATTATC |
| pBetI_EB | pBet_For_E | ATGAAGACGTGCTTTTCATGGATTCGTTACCAATTGAC | pBetI_Rev_B | ACGAAGACCTAGTAGCTAGCATTATATTGAACGTCCAA |
| pPhlF_GB | pPhlF_For_G | ATGAAGACGTTGCCTTCGTTACCAATTGACATGATACG | pPhlF_Rev_B | ACGAAGACCTAGTAACCTTAACGATACGGTACGTTTCG |
| pBM3R1_FB | pBM3R1_For_F | ATGAAGACGTCGCTTTCGTTACCAATTGACGGAATGAA | pBM3R1_Rev_B | ACGAAGACCTAGTAGCTAGCATTATCGGAATGAACGTT |
Primer Design for Fusion Proteins
| Device Name | Primer | Sequence |
|---|---|---|
| C0080_CI | C0080_Rev_I | ACGAAGACCTTAGACAACTTGACGGCTACATCATTCAC |
| C0040_CI | C0040_Rev_I | ACGAAGACCTTAGACAACTTGACGGCTACATCATTCAC |
| E0040m_ID | E0040m_For_I | ATGAAGACGTTCTAGAATGCGTAAAGGAGAAGAACTTTTC |
| E0030_ID | E0030_For_I | ATGAAGACGTTCTAGAATGGTGAGCAAGGGCGAGGAGCTG |
Flow Cytometry Data
| For the first tandem promoter flow cytometry experiment, we tested pBad-pTet-BCD2-E1010m-B0015 and pTet-pBad-BCD2-E1010m-B0015. We planned on inducing each promoter separately with it's corresponding small molecule (either arabinose or atc) and also planned on inducing both together. We followed the flow cytometry workflow. By growing the constructs in different concentrations of media, we hoped to see RFP fluorescence increase as the small molecule concentration increased. For each concentration, we also had a negative. We also ran controls including: J04B2RM, J04B2GM, COXGR, COXRG, and DH5alpha. |  |
| According to literature, the pTet-pBad construct has not previously functioned as suspected. This was possibly due to position-dependence interference [1]. Conversely, our flow cytometer data showed that pTet-pBad had a greater range of fluorescence than pBad-pTet. We will need to do further investigating to find out why this was the case. |  |
