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Team:Bordeaux/NotebookJune5
From 2014.igem.org
(Difference between revisions)
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<li class="d-detail">We obtain three ELPs strains streaked on LB medium | <li class="d-detail">We obtain three ELPs strains streaked on LB medium | ||
<ul class="strain-title"> | <ul class="strain-title"> | ||
| - | <li>A strain of ELP VPGXG-20<br> | + | <li><b>A strain of ELP VPGXG-20</b><br> |
Strain A : pUC19-New-Met20 in <a href='https://www.neb.com/products/c2992-neb-5-alpha-fiq-competent-e-coli-high-efficiency'>NEB 5-alpha F'Iq Competent E. coli</a> | Strain A : pUC19-New-Met20 in <a href='https://www.neb.com/products/c2992-neb-5-alpha-fiq-competent-e-coli-high-efficiency'>NEB 5-alpha F'Iq Competent E. coli</a> | ||
<ol class="strain"> <li> Used for cloning or others </li> | <ol class="strain"> <li> Used for cloning or others </li> | ||
<li> Amp<sup>R</sup> </li> | <li> Amp<sup>R</sup> </li> | ||
</ol></li> | </ol></li> | ||
| - | <li>A strain of ELP VPGXG-40<br> | + | <li><b>A strain of ELP VPGXG-40</b><br> |
Strain B : pUC19-New-Met40 in Stbl3 E.coli (The Stbl3 E. coli strain is derived from the HB101 E. coli strain and is recommended for use when cloning unstable inserts such as lentiviral DNA containing direct repeats) | Strain B : pUC19-New-Met40 in Stbl3 E.coli (The Stbl3 E. coli strain is derived from the HB101 E. coli strain and is recommended for use when cloning unstable inserts such as lentiviral DNA containing direct repeats) | ||
<ol class="strain"> <li> Used for cloning or others </li> | <ol class="strain"> <li> Used for cloning or others </li> | ||
<li> Amp<sup>R</sup> </li> | <li> Amp<sup>R</sup> </li> | ||
</ol></li> | </ol></li> | ||
| - | <li> A strain of ELP VPGXG-60<br> | + | <li><b> A strain of ELP VPGXG-60</b><br> |
Strain C : pUC19-GS60 in Stbl3 E.coli | Strain C : pUC19-GS60 in Stbl3 E.coli | ||
<ol class="strain"> <li> Used for cloning or others </li> | <ol class="strain"> <li> Used for cloning or others </li> | ||
| Line 46: | Line 46: | ||
<li class="d-detail">We use BL21 E.coli for the production </li><br> | <li class="d-detail">We use BL21 E.coli for the production </li><br> | ||
<li class="d-detail"> We obtain three ELP strains streaked on LB medium : | <li class="d-detail"> We obtain three ELP strains streaked on LB medium : | ||
| - | <ul class="strain-title"><li> A strains of ELP VPGXG-20<br> | + | <ul class="strain-title"><li> <b>A strains of ELP VPGXG-20 </b><br> |
| - | Pattern: MW-(VPGXG)<sub>20</sub><br> | + | <br>Pattern: MW-(VPGXG)<sub>20</sub><br> |
Number of amino acids : 102<br> | Number of amino acids : 102<br> | ||
Molecular weight : 8685.4 Da<br> | Molecular weight : 8685.4 Da<br> | ||
| - | Theoretical pI : 5.28<br> | + | Theoretical pI : 5.28<br><br> |
Strain A’ : pET44a-New-Met20 in BL21 E.coli | Strain A’ : pET44a-New-Met20 in BL21 E.coli | ||
<ol class="strain"> <li> Used for cloning or others </li> | <ol class="strain"> <li> Used for cloning or others </li> | ||
<li> Amp<sup>R</sup> </li> | <li> Amp<sup>R</sup> </li> | ||
</ol></li> | </ol></li> | ||
| - | <li>A strain of ELP VPGXG-40<br> | + | <li> <b>A strain of ELP VPGXG-40</b><br> |
| - | Pattern : MW-(VPGXG)<sub>40</sub><br> | + | <br>Pattern : MW-(VPGXG)<sub>40</sub><br> |
Number of amino acids : 202<br> | Number of amino acids : 202<br> | ||
Molecular weight : 17035.4 Da<br> | Molecular weight : 17035.4 Da<br> | ||
| - | Theoretical pI : 5.28<br> | + | Theoretical pI : 5.28<br><br> |
Strain B’ : pET44a-New-Met40 in BL21 E.coli | Strain B’ : pET44a-New-Met40 in BL21 E.coli | ||
<ol class="strain"> <li> Used for cloning or others </li> | <ol class="strain"> <li> Used for cloning or others </li> | ||
<li> Amp<sup>R</sup> </li> | <li> Amp<sup>R</sup> </li> | ||
</ol></li> | </ol></li> | ||
| - | <li>A strain of ELP VPGXG-60<br> | + | <li> <b>A strain of ELP VPGXG-60</b><br> |
| - | Pattern : MW-(VPGXG)<sub>60</sub><br> | + | <br>Pattern : MW-(VPGXG)<sub>60</sub><br> |
Number of amino acids : 303<br> | Number of amino acids : 303<br> | ||
Molecular weight : 25849.3 Da<br> | Molecular weight : 25849.3 Da<br> | ||
| - | Theoretical pI : 5.27<br> | + | Theoretical pI : 5.27<br><br> |
Strain C’ : pET44a-GS60 in BL21 E.coli | Strain C’ : pET44a-GS60 in BL21 E.coli | ||
Revision as of 20:31, 17 October 2014
5th June
Molecular biology
- We obtain three ELPs strains streaked on LB medium
- A strain of ELP VPGXG-20
Strain A : pUC19-New-Met20 in NEB 5-alpha F'Iq Competent E. coli- Used for cloning or others
- AmpR
- A strain of ELP VPGXG-40
Strain B : pUC19-New-Met40 in Stbl3 E.coli (The Stbl3 E. coli strain is derived from the HB101 E. coli strain and is recommended for use when cloning unstable inserts such as lentiviral DNA containing direct repeats)- Used for cloning or others
- AmpR
- A strain of ELP VPGXG-60
Strain C : pUC19-GS60 in Stbl3 E.coli- Used for cloning or others
- AmpR
- A strain of ELP VPGXG-20
- Inoculation of 3 LB medium for store
- Transformation by electroporation of our bacteria with low copy IGEM backbone pSB6A1(2014 Kit Plate 4, well 2L)
Production
- We use BL21 E.coli for the production
- We obtain three ELP strains streaked on LB medium :
- A strains of ELP VPGXG-20
Pattern: MW-(VPGXG)20
Number of amino acids : 102
Molecular weight : 8685.4 Da
Theoretical pI : 5.28
Strain A’ : pET44a-New-Met20 in BL21 E.coli- Used for cloning or others
- AmpR
- A strain of ELP VPGXG-40
Pattern : MW-(VPGXG)40
Number of amino acids : 202
Molecular weight : 17035.4 Da
Theoretical pI : 5.28
Strain B’ : pET44a-New-Met40 in BL21 E.coli- Used for cloning or others
- AmpR
- A strain of ELP VPGXG-60
Pattern : MW-(VPGXG)60
Number of amino acids : 303
Molecular weight : 25849.3 Da
Theoretical pI : 5.27
Strain C’ : pET44a-GS60 in BL21 E.coli- Used for cloning or others
- AmpR
- A strains of ELP VPGXG-20
