Team:Paris/August 6
From 2008.igem.org
(→Assay to quantify DNA) |
(→Assay to quantify DNA) |
||
Line 27: | Line 27: | ||
[[Image:Gel 080806.JPG|thumb|]] | [[Image:Gel 080806.JPG|thumb|]] | ||
- | Results : The MP123 plasmid is clearly visible but the PCR products (purified by QIAquick Gel Extraction) | + | Results : The MP123 plasmid is clearly visible but the PCR products (purified by QIAquick Gel Extraction) aren't. There might be a problem during the purification step using the QIAquick Gel Extraction. Whatever, we still go on with the digestion. |
- | + | Besides the MP123 sample was quantified using a Bio Photometer (Eppendorf). | |
{| Border="1" | {| Border="1" |
Revision as of 13:32, 6 August 2008
Plasmid extractionThe J61002 plasmid was extrated from overnight bacteria culture. The number of this plasmid extraction is MP123. This extraction has been conducted 3 times. Assay to quantify DNAIn order to quantify the DNA contained either in the Miniprep product of MP123 or in the PCR products of MP124, MP125, MP126 and MP127 previously purifed by QIAquick Gel Extraction yesterday, we carried out an electrophoresis assay in a 1,5% agarose gel. The ladder used was the 100 bp DNA ladder (New England Biolabs). We added 1 µL of 6X blue loading dye on 5 µL of each sample, which were then loaded on the gel.
Results : The MP123 plasmid is clearly visible but the PCR products (purified by QIAquick Gel Extraction) aren't. There might be a problem during the purification step using the QIAquick Gel Extraction. Whatever, we still go on with the digestion. Besides the MP123 sample was quantified using a Bio Photometer (Eppendorf).
DigestionFor each sample (MP123 and PCR products of MP124, MP125, MP126 and MP127 previously purified by QIAquick Gel Extraction):
The reaction was incubated 1h30 at 37°C, then 1h30 more after adding 1 µL of SpeI. |