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You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.

Contents 1 Methods and Protocols 1.1 Culture Medium 2 Agar Plate 2.1 LBampicillin-Agar 2.2 SCD-Agar 2.3 YEPDG418-Agar 3 Gel Electrophoresis 4 Kit 4.1 PCR Purification Kit 4.2 Gel Extraction Kit 4.3 Midi Plasmid Preparation Kit

Methods and Protocols

Culture Medium

Full Medium (YEPD) for Yeast
Yeast Extract	1 % (weight/volume)
Pepton	2 % (w/v)
Glucose	2 % (w/v)

Synthetic Complete Medium (SC) for Yeast
Yeast Nitrogen Base	0.17 % (w/v)
Ammoniumsulfate	0.5 % (w/v)
Amino Acid Mix*	50 ml/l
Histidin**	0.25 mM
Tryptophan**	0.19 mM
Leucin**	0.35 mM
Uracil**	0.44 mM

pH has to be regulated with KOH to pH=6.3
!* contains no His, Leu, Trp and Uracil
** addition of this components depents on the respective selection medium

SOC-Medium for Regeneration of transformed Escherichia coli`s after Electroporation
Trypton	2 % (w/v)
Yeast Extract	0.5 % (w/v)
NaCl	10 mM
KCl	2,5 mM
MgCl2	10 mM
MgSO4	10 mM
Glucose	20 mM

pH has to be regulated to pH=6.8-7.0

Full Medium (YEPD) for Yeast
Yeast Extract	1 % (weight/volume)
Trypton	0.5 % (w/v)
NaCl	0.5 % (w/v)

pH has to be regulated with NaOH to pH=7.5

Every cluture medium has to be autoclaved.

Agar Plate

LB_ampicillin-Agar

Add 2 % agar to LB-medium. After autoclaving and cooling-down to 60 °C steril ampicillin is added. Plates were poured.

SCD-Agar

Add 2 % agar to SCD-medium. After autoclaving and cooling-down steril amino acid solution is added. Dependent on the respective selective medium Histidin (0.25 mM), Trypthophan (0.19 mM), Uracil (0.44 mM) or Leucin (0.35 mM) is added.

YEPD_G418-Agar

Add 2 % agar to YEPD-medium. After autoclaving and cooling-down sterile G418 (final concentration 2g/l) is added.

Gel Electrophoresis

Agarose Gel (1x)
TAE puffer	1x
Agarose	1 % (w/v)

Solve agarose in TAE by boiling it. After cooling-down to 55-60 °C gel is poured.

TAE Puffer (50x)
EDTA	18,6 g
Tris	242g
Glacial Acetic Acid	57,2 ml
Purified Water	1000ml

pH has to be regulated with glacial acetic acid to pH=8.

Kit

PCR Purification Kit

Gel Extraction Kit

Midi Plasmid Preparation Kit