Difference between revisions of "Team:Paris Saclay/Notebook/July/1"
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{{Team:Paris_Saclay/notebook_header}} | {{Team:Paris_Saclay/notebook_header}} | ||
| − | = | + | =Friday 1<sup>st</sup> July= |
| − | + | ||
===Biobrick characterization=== | ===Biobrick characterization=== | ||
====Buffer preparations==== | ====Buffer preparations==== | ||
| − | '' | + | ''By Charlene and Caroline'' |
| + | The following buffers were prepared: | ||
{| class="wikitable" | {| class="wikitable" | ||
! Buffer Z (250mL) | ! Buffer Z (250mL) | ||
| − | ! Buffer | + | ! Buffer Luc (250mL) |
! Buffer STOP (15mL) | ! Buffer STOP (15mL) | ||
|- | |- | ||
| − | | | + | | |
| − | + | Na<sub>2</sub>HPO<sub>4</sub> – 3,125g | |
| − | + | ||
| − | + | NaH<sub>2</sub>PO<sub>4</sub>.H<sub>2</sub>O – 1,38g | |
| − | + | ||
| − | | Trisphosphate pH 7,8 1M – 6,25mL | + | KCl – 0,185g |
| − | + | ||
| − | + | MgSO<sub>4</sub> – 0,04g | |
| − | + | ||
| − | + | β-mercaptoethanol – 0,893mL | |
| − | + | ||
| − | + | | | |
| − | | | + | Trisphosphate pH 7,8 1M – 6,25mL |
| + | |||
| + | MgCl<sub>2</sub> 2M – 1mL | ||
| + | |||
| + | DTT 1M -0,25mL | ||
| + | |||
| + | EDTA 0,5M – 0,5mL | ||
| + | |||
| + | BSA – 0,25g | ||
| + | |||
| + | Glycerol 60% - 62,5mL | ||
| + | |||
| + | Triton 10% (100x) – 25mL | ||
| + | | | ||
| + | Na<sub>2</sub>CO<sub>3</sub> 1M – 0,78g | ||
|} | |} | ||
| + | |||
| + | ===Visualization=== | ||
| + | ====Migration of plasmids containing gBlocks digestion==== | ||
| + | ''By Léa and Alice'' | ||
| + | |||
| + | Digestion products from 30/06/16 were migrated on a gel (0.8% agarose) for 30 min at 100 V. | ||
| + | [[File:T--Paris_Saclay--160701_visualization_gBlocks_échelle.jpg|400px|thumb|right|pPS16_002, pPS16_003 and pPS16_005 Migration]] | ||
| + | [[File:T--Paris_Saclay--160701_visualization_gBlocks2_échelle.jpg#file|400px|thumb|right|pPS16_007, pPS16_008 and pPS16_009 Migration]] | ||
| + | |||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | ! Component | ||
| + | ! Volume (µL) | ||
| + | |- | ||
| + | | Digested DNA | ||
| + | | 10 | ||
| + | |- | ||
| + | | Loading buffer | ||
| + | | 2 | ||
| + | |} | ||
| + | |||
| + | |||
| + | For strains containing [[Team:Paris_Saclay/Notebook/June/28#pPS16_007|pPS16_007]] plasmid, according to the digestion products, 2 clones had the plasmid with the good insert. Indeed we could observe on the gel the two expected fragments of 2.7kb and 0.7kb. The same was observed for strains containing gBlocks 4.2 and GFP 1-9, respectively 3 and 2 clones were good. However, DNA concentration was too low. | ||
| + | For others strains, migration products did not get consistent results, probably due to errors during the extraction. | ||
| + | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 13:56, 9 October 2016
