G.tigrina Hox gene DthoxC insertion into prokaryote E.coli / (by UNIamCloning)/2011/10/25: Difference between revisions
(Autocreate 2011/10/25 Entry for G.tigrina_Hox_gene_DthoxC_insertion_into_prokaryote_E.coli_/_(by_UNIamCloning)) |
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== | ==Transformation of E.coli competent cells with resuspended GFP and ATF1 DNA== | ||
Today we located our GFP and ATF1 Biobrick parts in the proper plates and took samples from each year: (2011, 2010, 2009). We resuspended the DNA using 15ul H2O and transformed it into 40ul competent cells. We also created a positive control using a special plasmid that SHOULD, if the transformation went smoothly, grow nicely on the ampicillin plates, as well as a negative control that should NOT grow on the ampicillin plates - (lest the plates are messed up) - but SHOULD grow on the non-antibiotic plates - (lest the transformation killed the cells). | |||
After adding 950ul - (from the protocol... next time, only add 200ul!) - SOC NP to the tubes of resuspended DNA and competent cells, we allowed them to "shake" - (mix) - for 60 minutes. Then, we spread the GFP and ATF1 mixes and our two controls onto ampicillin plates, and our negative control and leftover competent cells onto non-antibiotic plates, and placed them into the oven at 37°C overnight. | |||
Revision as of 15:28, 25 October 2011
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Transformation of E.coli competent cells with resuspended GFP and ATF1 DNAToday we located our GFP and ATF1 Biobrick parts in the proper plates and took samples from each year: (2011, 2010, 2009). We resuspended the DNA using 15ul H2O and transformed it into 40ul competent cells. We also created a positive control using a special plasmid that SHOULD, if the transformation went smoothly, grow nicely on the ampicillin plates, as well as a negative control that should NOT grow on the ampicillin plates - (lest the plates are messed up) - but SHOULD grow on the non-antibiotic plates - (lest the transformation killed the cells). After adding 950ul - (from the protocol... next time, only add 200ul!) - SOC NP to the tubes of resuspended DNA and competent cells, we allowed them to "shake" - (mix) - for 60 minutes. Then, we spread the GFP and ATF1 mixes and our two controls onto ampicillin plates, and our negative control and leftover competent cells onto non-antibiotic plates, and placed them into the oven at 37°C overnight.
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