Materials, instruments and source of supplies

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Lys ozyme Si gma -Al dri ch, Ta ufki rchen

Mi l l ex® AP20 Syri nge Fi l ter, pore s i ze 20 µm Merck KGa A, Da rms ta dt

NEBNext® Sma l l RNA Li bra ry Prep Set New Engl a nd Bi ol a bs GmbH, Fra nkfurt NEB Bui l der Hi Fi DNA As s embl y Ma s ter Mi x New Engl a nd Bi ol a bs GmbH, Fra nkfurt

NTPs (ATP/GTP/CTP/UTP) Jena Bi os ci ence GmbH, Jena

Phenol /Chl oroform Mi x (a ci di c) Ambi on, Da rms ta dt

Phus i on DNA pol ymera s e Thermo Fi s her Sci enti fi c Ltd. & Co. KG, Bonn

QIAprep Spi n Mi ni prep Ki t Qi a gen GmbH, Hi l den

QIAqui ck Gel Extra cti on Ki t Qi a gen GmbH, Hi l den

QIAqui ck PCR Puri fi ca ti on Ki t Qi a gen GmbH, Hi l den

Qubi t ds DNA HS As s a y Ki t Thermo Fi s her Sci enti fi c Ltd. & Co. KG, Bonn Qubi t RNA HS As s a y Ki t Thermo Fi s her Sci enti fi c Ltd. & Co. KG, Bonn Qui ck-Loa d® 2-Log DNA La dder (0.1-10.0 kb) New Engl a nd Bi ol a bs GmbH, Fra nkfurt Qui ck-Loa d® pBR322 DNA-Ms pI Di ges t New Engl a nd Bi ol a bs GmbH, Fra nkfurt Res tri cti on endonucl ea s es New Engl a nd Bi ol a bs GmbH, Fra nkfurt

RNa s e If New Engl a nd Bi ol a bs GmbH, Fra nkfurt

RNa s e Inhi bi tor (muri ne) New Engl a nd Bi ol a bs GmbH, Fra nkfurt Roti ®-Nyl on pl us , pore s i ze 0.45 μm Roth GmbH, Ka rl s ruhe

Sodi um dodecyl s ul fa te (SDS) Roth GmbH, Ka rl s ruhe

Si gma PrepTM Spi n Col umn Ki t Si gma -Al dri ch, Ta ufki rchen

Sumo Protea s e Own producti on

SYBR Gol d® Nucl ei c a ci d s ta i n Thermo Fi s her Sci enti fi c Ltd. & Co. KG, Bonn

T4 DNA Li ga s e New Engl a nd Bi ol a bs GmbH, Fra nkfurt

T4 Pol ynucl eoti de Ki na s e New Engl a nd Bi ol a bs GmbH, Fra nkfurt

T7 RNA Pol ymera s e New Engl a nd Bi ol a bs GmbH, Fra nkfurt

Tetra methyl ethyl endi a mi ne (TEMED) Si gma -Al dri ch, Ta ufki rchen ULTRAhyb-Ol i go hybri di za ti on buffer Ambi on, Da rms ta dt

Wha tma n GB 004, 3MM Schl ei cher & Schuel l GmbH, Da s s el

X-Ra y CEA RP New fi l m s creen CEA GmbH, Ha mburg

Zel l uTra ns Di a l ys i s tubes (6,000-8,000 MWCO) Roth GmbH, Ka rl s ruhe

The general chemicals and reagents that are not listed above were purchased from AppliChem GmbH (Darmstadt), BioRad Laboratories GmbH (München), Biozym GmbH (HessischOldendorf), Difco Laboratories GmbH (Augsburg), Invitrogen (Karlsruhe), Merck KGaA (Darmstadt), Roche GmbH (Mannheim), Roth GmbH (Karlsruhe), VWR International (Darmstadt), SERVA GmbH (Heidelberg) and Sigma-Aldrich Co. (Taufkirchen).

107 4.1.2 Instruments

Table 4.2: List of Instruments used in this study

Ins trument Model and Company

Aga ros e gel electrophoresis Cha mbers a nd Ca sting trays produced by company technician Philipps-University Ma rburg

Power s upply Consort E835; MS La borgeräte, Di elheim

Aqua bidest, water s ystem PURELAB Pl us, ELGA La bWater, Celle Autocl ave 5075 EL, Tuttna uer Europe B.V., Breda, NL

Bi oanalyzer Agi l ent 2100 Bi oanalyze r, Agilent, Santa Clara, CA, USA

Centri fuges Centri fuge 5424, Eppendorf AG, Hamburg; Si gma 3-30K, Si gma Laborzentrifugen GmbH, Os terode a m Harz; Sorvall Lynx 4000, Thermo Fisher Scientific Ltd. & Co. KG, Bonn Chroma tography columns Hi s Trap HP 1 ml a nd 5 ml , StrepTrap HP 5 ml , HiLoad® 16/600 Superdex® 200 pg,

Superose® 6 10/300 GL, Mono Q 5/50 GL a nion exchange chromatography column; GE Hea lthcare Europe GmbH, Freiburg

Fl ow cytometer BD LSRFortessa, BD Biosciences, Heidelberg

FPLC ÄKTA puri fier™ 10: Pump P-900, Moni tor UV-900, Monitor UPC-900, Va lve INV-907, Mi xer M-925; ÄKTA pure L1: Pump P9 Cpl , Mi xer M9, Injection valve V9-Inj, UV-Monitor U9-L Cpl ; GE Hea lthcare Europe GmbH, Freiburg

Denaturing polyacrylamide gel el ectrophoresis

PROTEAN II El ectrophoresis Chamber, BioRad La boratories GmbH, Munich Hybri dization oven Hybri d Shake 'n' Stack, Thermo Fisher Scientific Ltd. & Co. KG, Bonn

Incubators KB53, Bi nder GmbH, Tuttlingen

Ma gnetic stirrer IKA® RCT Sta ndard, IKA®-Werke GmbH & Co. KG, Sta ufen

Ma gnetic Separation Rack 2-Tube Ma gnetic Separation Rack; New England Biolabs GmbH, Frankfurt

Mi cros cope Axi oplan 2, Ca rl Zeiss Mi croscopy GmbH, Göttingen; CoolSnap HQ camera, Visitron Sys tems GmbH, Puchheim; FluoArc HBO Lamp, Ca rl Zeiss Mi croscopy Gmb H, Göttingen Mi crofl uidizer Mi crofl uidics LM10, Sys mex Deutschland GmbH, Norderstedt.

Mi nIon Oxford Na nopore Technologies, UK

Mi ni Seq Il l umina , Inc. USA

Na nodrop Na noDrop® ND-1000 Spectrometer, Thermo Fisher Scientific Ltd. & Co. KG, Bonn PCR-Cycl er C1000TM Thermal Cycl er, Bi oRad La boratories GmbH, Munich

Peri s taltic pump Peri s taltic Pump P-1, GE Healthcare Europe GmbH, Freiburg

pH-meter INOLAB pH l evel 1, WTW, Weilheim

Phos phorimager Storm 840 phos phorimager, Molecular Dynamics, GE Healthcare Europe GmbH, Frei burg

Qubi t Fluorometer Qubi t 2.0, Thermo Fisher Sci entific Ltd. & Co. KG, Bonn Rocker Gyrorocker SSL3, Si gma-Aldrich, Taufkirchen

Sci ntillation counter Beckmann LS 6500, Beckman Coulter GmbH, Krefeld

Semi -dry tra nsfer cell Tra ns -Blot® SD Semi-Dry Tra nsfer Cell, BioRad Laboratories GmbH, Munich SDS pol yacrylamide gel

el ectrophoresis

Mi ni -PROTEAN Tetra Cell, Bio-Rad Laboratories GmbH, Munich; Power s upply PowerPac Ba s ic, Bio-Rad La boratories GmbH, Munich

Soni cator Bra nson Sonifier 250, Bra nson Ultrasonicx, Danbury, CT, USA Spectrophotometer Ul tros pec 3000 pro, GE Healthcare Europe GmbH, Freiburg Thermomixer Thermomixer Comfort 5350, Eppendorf AG, Hamburg Thermoshaker HT Thermotron, Infors AG, Bottmingen, Switzerland UV-Cros slinker UV Stra ta linker® 1800, Stra tagene, La Jolla, USA UV-Tra nsilluminator Bi oDocd-IT s ystem, UVP, Upland, CA, USA

Vortex Mi xer Vortex Genie 2, Sci entific Industries, Bohemia, NY, USA

4.1.3 Buffers and solutions

Buffers, solutions and media are mentioned in the chapter of the respective method. All media, solutions and buffers were, if necessary, autoclaved for 20 min at 121°C prior to usage.

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