- Backup comparator circuit mini preps and digest with Eco R1 and Pst1 (overnight)
+
. As a backup, the running some of the running culture of the Comparator circuit 1 and 2 transformed cells were mini-prepped. After nanodropping, the concentration was found to be.... Following this an overnight double restriction digest with ''Eco''R1 and ''Pst''1 was set up. These were set up using: 0.2 BSA, 2 Buffer H, 0.5 EcoR1 and 0.5 Pst1. As to the DNA, the following amounts were added.
-
- BM minipreped (didn't work so re-inoculated)
+
. Following a low concentration reading on Friday, B-M was miniprepped again. However, this produced no DNA when nanodropped. Therefore, B-M was reinoculated into culture.
-
- Digest MB samples with Spe1 and Pst1, cut out and purify
-
- CFP and RFP digested with PstI and XbaI (cut out and gell purified)
+
. Digest MB samples with Spe1 and Pst1, cut out and purify
-
- Ligate MB2, MB3,MB5,MB9,MB10 with RFP/CFP (overnight)
+
. CFP and RFP digested with PstI and XbaI (cut out and gell purified)
-
- Ligate Comparator insert with PSB1C3 (overnight)
+
. Ligate MB2, MB3,MB5,MB9,MB10 with RFP/CFP (overnight)
+
+
. Ligate Comparator insert with PSB1C3 (overnight)
. As a backup, the running some of the running culture of the Comparator circuit 1 and 2 transformed cells were mini-prepped. After nanodropping, the concentration was found to be.... Following this an overnight double restriction digest with EcoR1 and Pst1 was set up. These were set up using: 0.2 BSA, 2 Buffer H, 0.5 EcoR1 and 0.5 Pst1. As to the DNA, the following amounts were added.
. Following a low concentration reading on Friday, B-M was miniprepped again. However, this produced no DNA when nanodropped. Therefore, B-M was reinoculated into culture.
. Digest MB samples with Spe1 and Pst1, cut out and purify
. CFP and RFP digested with PstI and XbaI (cut out and gell purified)
. Ligate MB2, MB3,MB5,MB9,MB10 with RFP/CFP (overnight)
. Ligate Comparator insert with PSB1C3 (overnight)
"
