{"date" : "2013-07-09","author" : "emil","title" : " Purification of Inocula (05/7) and re-ligation and re-transformation ","content" : "Unfortunately only one of the inocula of 05/7 has succeded and shows an incorrect red color(RFP the previous insert) so I decided to purify and quantfy only the 2 attempt to amplify 024.I have purified them following the purification protocol these are the results of the quantification.

BBa_K823024+BBa_E0840(1:1) 2267 ng/µl
BBa_K823024+BBa_E0840(1:1) 3248.6 ng/µl
Afterwards I screened 800 ng of the samples following the screening protocol with EcoR1 HF and Pst1, these are the results of the gel:
Gel order
BBa_K823024+BBa_E0840(1:1) b1
BBa_K823024+BBa_E0840(1:1) 2
Ladder 1kb Fermentas3
No one of the sample has succeded there are only the bands of the backbone and ther aren't visible inserts of any kind, so I decided to re-try the ligation with two couples of 026 and GFP(026a=36.3 ng/µl,026b=37.6ng/µl,GFPa=15.9ng/µl,GFPb015.8ng/µl,I have done it following the ligation protocol (I have only changed the amaunt of plasmid:300 ng).Afterwards I have transformed and plated on ampicillin agar 8 NEB10β (2 ctrl,2 1:1, 2 1:2, 2 1:3).I have also amplified the 024(2 inocula).","tags" : "K832024-K823026-E0840"}