|
|
(15 intermediate revisions by the same user not shown) |
Line 1: |
Line 1: |
| =Pseudomonas aeruginosa mer operon cloning 3=
| | |
| ==Preparation==
| |
| What can I do in order to make sure the cloning works this time?
| |
| *Check concentrations of the chromatin DNA
| |
| *Do a dilution series over three orders of magnitude of the template DNA
| |
| ==PCR set-up of the operon==
| |
| #Master mix
| |
| ##Template DNA equiv.
| |
| ##ddH<sub>2</sub>O to 50uL
| |
| #Thermal profile
| |
| ##92 deg C - 5min
| |
| ##92 deg C - 30s
| |
| ##50 deg C - 45s
| |
| ##72 deg C - 120s
| |
| ##4 deg C - indefinite
| |
| ===Pipetting scheme===
| |
| {| border="1"
| |
| ! Column heading 1 !! Column heading 2 !! Column heading 3
| |
| |-
| |
| ! Row heading 1
| |
| | Cell 2 || Cell 3
| |
| |-
| |
| ! Row heading A
| |
| |Cell B
| |
| |Cell C
| |
| |}
| |
| ==Gel image==
| |
| 1% agarose TAE gel, ran @ 100V for 60min
| |