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Team:Bielefeld-Germany/Labjournal/week12
From 2012.igem.org
Contents |
Labjournal
Week1 Week2 Week3 Week4 Week5 Week6 Week7 Week8 Week9 Week10 Week11 Week12 Week13 Week14 Week15 Week16 Week17 Week18
Week 12 (07/16 - 07/22/12)
Monday July 16th
- Team A. thaliana laccase: Our plants had a great time during the last weeks in the climate chamber. So today it was time for them to donate their seeds for RNA isolation, cDNA synthesis and a PCR (check protocols). We ran an additional sample with actin primers as a positive control. However both samples did not show any bands. Maybe the high salt concentration in our sample was responsible or the laccase concentration in the 1:10 deluted cDNA was too low. We will do some washing and try again.
Tuesday July 17th
- Team Cultivation/Purification: Second cultivation
- Team Site Directed Mutagenesis: Made Clone Manager files for Trametes versicolor sequences
Wednesday July 18th
- Team Site Directed Mutagenesis: Made Primer-Mixes. Got everything ready for Lab work.
Thursday July 19th
- Team Site Directed Mutagenesis:
- Team Cellulose Binding Domain: -
- Got info about Carbohydrate binding domain X2 (which is a more common domain in the organisms we handle than a Cellulose Binding Domain.)
- Nucleotid BLASTed BBa_K392014 (Cellulose-binding motif from C. josui Xyn10A gene) -> 1526-2608 of AB041993.1 (Clostridium josui xynA gene for xylanase A) - Port BLASTed AB041993.1 -> got two cellulose binding sites: - pfam02018: CBM_4_9 (Carbohydrate binding domain) from (193) AETFKTITFED to SASS (332) - corresponds to 1088 - 1507 in Gene and is not in CDS of BBa_K392014 - cd09619: CBM9_like_4 (DOMON-like type 9 carbohydrate binding module) From (716) TPVIDGTIDSVWS to TGYQDTSVYG (887) - corresponds to 2657 - 3172 in Gene an is not in CDS of BBa_K392014 - Given sequence of BBa_K392014 (1589-2590) corresponds to the sequence of the Glycosyl hydrolase 1589-2590 and not one of the binding motifs
Friday July 20th
- Team Modeling: We need a contact to a clarification plant to get information about clarification plant itself and perhaps to proof our cleaner with real probes. therefor we are calling Mr. Bülter form the clarification plant Schloß Holte (near Bielefeld) and he invited us to present our projekt.
