Fri 19 Aug 2011

Vessa

To Do List

• Transform 2 construct
• Digest RBS-RFP 6 plasmid with EP and XP
• Sequences data analysis!!

12 variants digestions
22.01 - 90 ng/ul
22.02 - 70 ng/u
22.03 - 53 ng/ul
22.04 - 53 ng/ul
22.05 - 72 ng/ul
22.06 - 32 ng/ul
22.07 - 66 ng/ul
22.08 - 47 ng/ul
22.09 - 39 ng/ul
22.10 - 43 ng/ul
22.11 - 35 ng/ul
22.12E - 152 ng/ul
22.12F - 156 ng/ul

Vector
18 ul MQ
3 ul 10x buffer
1 ulSpeI
1 ul Psti
1 ul Fast alkaline phosphatase
6 ul vector

Insert
196 ul MQ
28 ul 10 buffer
14 ul XbaI
14 ul PstI
28 ul Insert

Ligation @reaction

2ul 10X buffer
1 ul T4 DNA ligase
7 ul Vector
10 ul Insert

Joyce

Plasmid prep of PhybB-taRNA (colonies 1.1 and 4.1), PBAD-RBS-GFP-DT (colonies 7 and 8) and PhybB-RBS-GFP-DT in TOP10 cells
(colonies 2 and 3)
Measurements with ND1000 nanodrop showed: DNA concentrations between 33ng/microliter to 70 ng/microliter
PhybB-taRNA samples and PBAD-RBS-GFP-DT samples were send for sequencing and glycerolstocks were made (also from PhybB-RBS-GFP-
DT in TOP10 cells)

Stagewerkplan was made
Sequencing results were checked: sequencing did not go well itself, so I have to wait for sequencing results of the plasmids
which were isolated today. If this is alright, the others were also alright.

Stageforms had to be filled in

All overnight cultures for flow cytometry grew! E.coli DH5alpha needs to be grown in M9 minimal medium with cas amino acids!
Measurements with the Flow cytometer spread over a few hours. Start: 13:00 h