Difference between revisions of "Team:IIT-Madras/Proof"
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== RIBOSON == | == RIBOSON == | ||
<p>'''Trigger RNA''': A part of RNA molecule, which codes for [http://parts.igem.org/Part:BBa_E1010 RFP] protein.<br> | <p>'''Trigger RNA''': A part of RNA molecule, which codes for [http://parts.igem.org/Part:BBa_E1010 RFP] protein.<br> | ||
| − | Switch RNA: Switch RNA was designed to repress the translation process by masking the RBS placed upstream of GFP protein coding part. After interacting with the RNA molecules of RFP gene, RIBOSON would allow the RBS to open up to bind to ribosomal machinery for protein synthesis to take place.</p> | + | '''Switch RNA''': '''Switch RNA''' was designed to repress the translation process by masking the RBS placed upstream of GFP protein coding part. After interacting with the RNA molecules of RFP gene, <span class="riboson">RIBOSON</span> would allow the RBS to open up to bind to ribosomal machinery for protein synthesis to take place.</p> |
| − | <p>We performed our RIBOSON experiment in <i>E. coli</i> BL21(DE3). We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956019 BBa_K1956019], which has | + | <p>We performed our <span class="riboson">RIBOSON</span> experiment in <i>E. coli</i> BL21(DE3). We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956019 BBa_K1956019], which has <span class="riboson">RIBOSON</span>_{rfp} switch under [http://parts.igem.org/Part:BBa_J23100 J23100] constitutive promoter, upstream of [http://parts.igem.org/Part:BBa_E0040 GFP] coding part and RFP generator under [http://parts.igem.org/Part:BBa_R0011 lacI+pL] promoter.</p> |
<p> We grew the cells in LB broth media + 40mM Glucose. After, 16 hrs of growth, we made two types of secondary culture: 1. Cells + LB broth + 40mM Glucose and 2. Cells + LB broth + 1mM IPTG.</p> | <p> We grew the cells in LB broth media + 40mM Glucose. After, 16 hrs of growth, we made two types of secondary culture: 1. Cells + LB broth + 40mM Glucose and 2. Cells + LB broth + 1mM IPTG.</p> | ||
<p>Positive Control: Device [http://parts.igem.org/Part:BBa_K1956015 K1956015], which produces GFP and RFP proteins.<br> | <p>Positive Control: Device [http://parts.igem.org/Part:BBa_K1956015 K1956015], which produces GFP and RFP proteins.<br> | ||
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== RIBOSOFF == | == RIBOSOFF == | ||
<p>'''Trigger RNA''': A part of RNA molecule, which codes for lacI protein.<br> | <p>'''Trigger RNA''': A part of RNA molecule, which codes for lacI protein.<br> | ||
| − | Switch RNA: Switch RNA was designed to activate the translation process by allowing the ribosomal machinery to bind to RBS, placed upstream of GFP protein coding part. After interacting with the RNA molecules of lacI gene, RIBOSOFF changes it's confirmation to mask the RBS and represses the translation process.</p> | + | '''Switch RNA''': '''Switch RNA''' was designed to activate the translation process by allowing the ribosomal machinery to bind to RBS, placed upstream of GFP protein coding part. After interacting with the RNA molecules of lacI gene, <span class="ribosoff">RIBOSOFF</span> changes it's confirmation to mask the RBS and represses the translation process.</p> |
| − | <p>We performed our RIBOSOFF experiment in <i>E. coli</i> DH5alpha. We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020], which has | + | <p>We performed our <span class="ribosoff">RIBOSOFF</span> experiment in <i>E. coli</i> DH5alpha. We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020], which has <span class="ribosoff">RIBOSOFF</span>_{lacI} switch under [http://parts.igem.org/Part:BBa_J23100 J23106] constitutive promoter, upstream of [http://parts.igem.org/Part:BBa_E0040 GFP] coding part and RFP generator under [http://parts.igem.org/Part:BBa_R0011 lacI+pL] promoter.</p> |
<p> We prepared two types of cells: 1. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] in pSB1A2 + [http://parts.igem.org/Part:BBa_K1956034 BBa_K1956034] in pSB1K3, 2. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] only. <br> | <p> We prepared two types of cells: 1. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] in pSB1A2 + [http://parts.igem.org/Part:BBa_K1956034 BBa_K1956034] in pSB1K3, 2. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] only. <br> | ||
Revision as of 17:51, 19 October 2016
Noise in Devices
RIBOS
RIBOSON
Trigger RNA: A part of RNA molecule, which codes for [http://parts.igem.org/Part:BBa_E1010 RFP] protein.
Switch RNA: Switch RNA was designed to repress the translation process by masking the RBS placed upstream of GFP protein coding part. After interacting with the RNA molecules of RFP gene, RIBOSON would allow the RBS to open up to bind to ribosomal machinery for protein synthesis to take place.
We performed our RIBOSON experiment in E. coli BL21(DE3). We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956019 BBa_K1956019], which has RIBOSON_{rfp} switch under [http://parts.igem.org/Part:BBa_J23100 J23100] constitutive promoter, upstream of [http://parts.igem.org/Part:BBa_E0040 GFP] coding part and RFP generator under [http://parts.igem.org/Part:BBa_R0011 lacI+pL] promoter.
We grew the cells in LB broth media + 40mM Glucose. After, 16 hrs of growth, we made two types of secondary culture: 1. Cells + LB broth + 40mM Glucose and 2. Cells + LB broth + 1mM IPTG.
Positive Control: Device [http://parts.igem.org/Part:BBa_K1956015 K1956015], which produces GFP and RFP proteins.
Negative Control: Device [http://parts.igem.org/Part:BBa_K1956034 K1956034], which produces lacI mRNA molecules.
RIBOSOFF
Trigger RNA: A part of RNA molecule, which codes for lacI protein.
Switch RNA: Switch RNA was designed to activate the translation process by allowing the ribosomal machinery to bind to RBS, placed upstream of GFP protein coding part. After interacting with the RNA molecules of lacI gene, RIBOSOFF changes it's confirmation to mask the RBS and represses the translation process.
We performed our RIBOSOFF experiment in E. coli DH5alpha. We transformed our cloned device [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020], which has RIBOSOFF_{lacI} switch under [http://parts.igem.org/Part:BBa_J23100 J23106] constitutive promoter, upstream of [http://parts.igem.org/Part:BBa_E0040 GFP] coding part and RFP generator under [http://parts.igem.org/Part:BBa_R0011 lacI+pL] promoter.
We prepared two types of cells: 1. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] in pSB1A2 + [http://parts.igem.org/Part:BBa_K1956034 BBa_K1956034] in pSB1K3, 2. [http://parts.igem.org/Part:BBa_K1956020 BBa_K1956020] only.
We grew the cells in LB broth media. After, 16 hrs of growth, we inoculated secondary cultures and collect samples at 12 hour of growth.
Positive Control: Device [http://parts.igem.org/Part:BBa_K19560015 K19560015], which produces GFP and RFP proteins.
Negative Control: Device [http://parts.igem.org/Part:BBa_K1956034 K1956034], which produces lacI mRNA molecules.
