Team:UNITN-Trento/Notebook/Labposts/07/15

From 2013.igem.org

{"date" : "2013-07-04","author" : "emil","title" : " Re-Inocula of the plate with the product of ligation and purification of the first plates ","content" : "I purified the inocula done the day before following the purification protocol.Then I quantified the results with the following results:

Quantification
SampleQuantification
K823026+E0840 1:3 A409.4ng/µl
K823026+E0840 1:3 B401.0ng/µl
K823026+E0840 1:1 C733.4ng/µl
K823026+E0840 1:1 D489.3ng/µl
K823024+E0840 1:1 E(succesful)281ng/µl
K823024+E0840 1:3 F341.1ng/µl
K823024+E0840 1:3 G220.3ng/µl
K823024+E0840 1:1 error129.3ng/µl
Then I screened the purified products following the digestion protocol.These are the results of the gel(1%):
Gel order
SampleWell
Ladder 1kb Fermentas1
BBa_K823026+BBa_E0840 A2
BBa_K823026+BBa_E0840 B3
BBa_K823026+BBa_E0840 C4
BBa_K823026+BBa_E0840 D5
BBa_K823024+BBa_E0840 E(the only succesful)6
BBa_K823024+BBa_E0840 F7
BBa_K823024+BBa_E0840 G9
As we can see ther are two bands for each well(like expected after the digestion) but only the lower band of the 6 well has the GFP(920 bp) as insert, the other have only the RFP witha promoter.So I decided to repeat the digestion of GFP(an old sample 42.6 ng/µl digested completely) and the same sample of the two backbone exactly like in the previous days.I did the inocula of the new transformation and also of two colonies of the plate that I tooK the E sample from(2 E sample(1:1 024+0840),1 1:1 024+0840,2 1:3 024+0840,2 1:1 026+0840, 2 1:3 026+0840).","tags" : "K832024-K823026-E0840"}