Team:TU Munich/Notebook/Labjournal

(Difference between revisions)

Revision as of 16:20, 23 June 2012

Investigator: Ingmar, Volker Aim of the experiment: Plasmid amplification

Operation Sequence:

melting of 100 µl Ca-competent E.coli XL1-Blue cells
addition of 1 µl of the Plasmid pKS2µHyg-PAL-4Cl-CHS
incubation for 30 min on ice
heat shock for 5 min at 37 °C
transfer of cells to 1 ml LB-medium without antibiotics and incubate at 37°C and 180 rpm for 30 min
plate 100 µl on Amp-LB-plate
sediment the leftover in a centrifuge(30 - 60 sec, 13 000 rpm) and resuspend the sediment in 100 µl LB-medium and plate it as well on Amp-LB-plate

Investigator: Ingmar, Volker Aim of the experiment: Generation of an RFC 25 compatible version of the pYes2 Vector.

volume	reagent
2.5 µl	10x Pfu Ultra II buffer
4 µl	Plasmid P7 pYes2_RFC25 MCS template
0.5 µl	1:10 dilution of O38 (10 pmol/µL)
0.5 µl	1:10 dilution of O39 ((10 pmol/µL)
17 µl	ddH2O
0.5 µl	dNTP mix
0.5 µl	Pfu Turbo DNA polymerase (2.5 U / µl)

@@ Line 5: / Line 5: @@
 Aim of the experiment: Plasmid amplification
+Operation Sequence:
+* melting of 100 µl Ca-competent  ''E.coli'' XL1-Blue cells
+* addition of 1 µl of the Plasmid pKS2µHyg-PAL-4Cl-CHS
+* incubation for 30 min on ice
+* heat shock for 5 min at 37 °C
+* transfer of cells to 1 ml LB-medium without antibiotics and incubate at 37°C and 180 rpm for 30 min
+* plate 100 µl on Amp-LB-plate
+* sediment the leftover in a centrifuge(30 - 60 sec, 13 000 rpm) and resuspend the sediment in 100 µl LB-medium and plate it as well on Amp-LB-plate
 ==Saturday, 23th June==