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Team:Warsaw/Calendar-Main/5 September 2009
From 2009.igem.org
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Results: | Results: | ||
* I propably took the wrong genome DNA sample for the previous reaction, there are products but none of them has the correct length. | * I propably took the wrong genome DNA sample for the previous reaction, there are products but none of them has the correct length. | ||
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| + | <h3><div style="text-align: center;">Assembly of endosomal detection operon</div></h3> | ||
| + | <h4>Marcin</h4> | ||
| + | Task 1: | ||
| + | * Prepare the bacterial cultures for isolation of following constructs: | ||
| + | ** [http://partsregistry.org/Part:pSB1A3<span style="color: black">pSB1A3</span>] plasmid containing cro coding sequence | ||
| + | ** [http://partsregistry.org/Part:pSB1A3<span style="color: black">pSB1A3</span>] plasmid containing p53 coding sequence | ||
| + | Task 2: | ||
| + | * Isolate the plasmids containing previously described constructs | ||
| + | Methods: | ||
| + | Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described[http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf" here] | ||
| + | Task 3: | ||
| + | * Digest previously isolated plasmids and verify the correctness of the ligation: | ||
| + | Methods: | ||
| + | * Reaction mixture composition: | ||
| + | <pre> | ||
| + | 2 μl purified plasmid DNA product | ||
| + | 0.5 μl XbaI (Fermentas) | ||
| + | 0.5 μl PstI (Fermentas) | ||
| + | 2 μl Buffer Tango (Fermentas) | ||
| + | 15 μl MQ water | ||
| + | </pre> | ||
| + | * Digest was performed 90 minutes | ||
| + | Results: | ||
| + | ====Comments:==== | ||
| + | * All samples of cro CDS on [http://partsregistry.org/Part:pSB1A3<span style="color: black">pSB1A3</span>] are only empty plasmids | ||
| + | * Samples of Bax CDS have biobricks but I am unable to determine whether this biobricks is Bax CDS or [http://partsregistry.org/Part:BBa_E0010<span style="color: black">BBa_E0010</span>] due to contamination with the latter one. It is obligated to prepare another digestion to reveal the identity of the insert | ||
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Revision as of 00:27, 7 September 2009
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Contents |
Acquiring the internaline A gene from Listeria monocytogenes st. EDG-e genome
Jarek
Tasks:
- Electrophoretical segregation of PCR products from previous day in 0,8% agarosis gel
- Preparation of another PCR reaction with L.monocytogenes genome DNA
Results:
- I propably took the wrong genome DNA sample for the previous reaction, there are products but none of them has the correct length.
Assembly of endosomal detection operon
Marcin
Task 1:
- Prepare the bacterial cultures for isolation of following constructs:
Task 2:
- Isolate the plasmids containing previously described constructs
Methods: Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described" here Task 3:
- Digest previously isolated plasmids and verify the correctness of the ligation:
Methods:
- Reaction mixture composition:
2 μl purified plasmid DNA product 0.5 μl XbaI (Fermentas) 0.5 μl PstI (Fermentas) 2 μl Buffer Tango (Fermentas) 15 μl MQ water
- Digest was performed 90 minutes
Results:
Comments:
- All samples of cro CDS on pSB1A3 are only empty plasmids
- Samples of Bax CDS have biobricks but I am unable to determine whether this biobricks is Bax CDS or BBa_E0010 due to contamination with the latter one. It is obligated to prepare another digestion to reveal the identity of the insert
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