Team:TU Munich/Notebook/Labjournal

(Difference between revisions)

Revision as of 16:25, 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 an 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 an 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)

Revision as of 16:23, 23 June 2012 (view source) VolkerMorath (Talk \| contribs) (→Transformation of E.coli XL1-Blue with pKS2µHyg-PAL-4Cl-CHS) ← Older edit		Revision as of 16:25, 23 June 2012 (view source) VolkerMorath (Talk \| contribs) (→Transformation of E.coli XL1-Blue with pKS2µHyg-PAL-4Cl-CHS) Newer edit →
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	'''Investigator: Ingmar, Volker'''		'''Investigator: Ingmar, Volker'''
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	Aim of the experiment: Plasmid amplification		Aim of the experiment: Plasmid amplification