EPF-Lausanne/14 August 2008

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(Difference between revisions)
 
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Yesterday we found out that F1610 has a size of 300bp, instead of 800bp. So there are two hypotheses: 1) The mini-prep is bad or 2) the biobrick from IGEM is bad.
Yesterday we found out that F1610 has a size of 300bp, instead of 800bp. So there are two hypotheses: 1) The mini-prep is bad or 2) the biobrick from IGEM is bad.
-
So we do another digestion serie with EcoRI and SpeI. We do with different mini-prep of F1610 and the last mini-prep of E1010 that work(for positiv control).
+
So we do another digestion series with EcoRI and SpeI. We do it with different mini-preps of F1610 and with the last mini-prep of E1010 that worked (as a positive control).
-
Some mini-prep from F1610 are too small, there isn't enough liquid so we do PCR with these (we use E1010 from last mini-prep as a positiv control).
+
Some mini-preps of F1610 haven't enough liquid left, so we do a PCR with these (we use E1010 from the last mini-prep as a positive control).
-
We do in same time PCR of all fragments that ligated E1010/F1610 to see if there is a fragment of 1500 bp.
+
At the same time we do a PCR of all plasmids that should have E1010/F1610 ligated to see if there is a fragment of 1500 bp.
-
===Resultat of Gel===
+
===Result of Gel===
All mini-preps of F1610 have a fragment of 300 bp. So the biobrick from IGEM doesn't work.
All mini-preps of F1610 have a fragment of 300 bp. So the biobrick from IGEM doesn't work.
-
All ligated fragments E1010/F1610 aren't ligate.
+
No ligated fragment E1010/F1610 is visible.
-
(see Gel picture on page 75 in notebook of lab)
+
(see Gel picture on page 75 in lab notebook)
==Test of specific primer of BBs==
==Test of specific primer of BBs==
-
We do a PCR to see if specific primers that Alexandre commanded work with biobrick.
+
We do a PCR to see if the specific primers that Alexandre ordered work with our biobricks.
-
We do gel tomorrow.
+
We will do the gel tomorrow.

Latest revision as of 09:38, 19 August 2008

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Contents

Molecular biology

We do a mini-prep of E1010, "F1610" and P0140

E1010/F1610

Yesterday we found out that F1610 has a size of 300bp, instead of 800bp. So there are two hypotheses: 1) The mini-prep is bad or 2) the biobrick from IGEM is bad.

So we do another digestion series with EcoRI and SpeI. We do it with different mini-preps of F1610 and with the last mini-prep of E1010 that worked (as a positive control).

Some mini-preps of F1610 haven't enough liquid left, so we do a PCR with these (we use E1010 from the last mini-prep as a positive control).

At the same time we do a PCR of all plasmids that should have E1010/F1610 ligated to see if there is a fragment of 1500 bp.

Result of Gel

All mini-preps of F1610 have a fragment of 300 bp. So the biobrick from IGEM doesn't work. No ligated fragment E1010/F1610 is visible.

(see Gel picture on page 75 in lab notebook)

Test of specific primer of BBs

We do a PCR to see if the specific primers that Alexandre ordered work with our biobricks. We will do the gel tomorrow.

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