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:
QuantificationSample | Quantification |
---|
K823026+E0840 1:3 A | 409.4ng/µl |
K823026+E0840 1:3 B | 401.0ng/µl |
K823026+E0840 1:1 C | 733.4ng/µl |
K823026+E0840 1:1 D | 489.3ng/µl |
K823024+E0840 1:1 E(succesful) | 281ng/µl |
K823024+E0840 1:3 F | 341.1ng/µl |
K823024+E0840 1:3 G | 220.3ng/µl |
K823024+E0840 1:1 error | 129.3ng/µl |
Then I screened the purified products following the
digestion protocol.These are the results of the gel(1%):
Gel orderSample | Well |
---|
Ladder 1kb Fermentas | 1 |
BBa_K823026+BBa_E0840 A | 2 |
BBa_K823026+BBa_E0840 B | 3 |
BBa_K823026+BBa_E0840 C | 4 |
BBa_K823026+BBa_E0840 D | 5 |
BBa_K823024+BBa_E0840 E(the only succesful) | 6 |
BBa_K823024+BBa_E0840 F | 7 |
BBa_K823024+BBa_E0840 G | 9 |
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"}