Main public logs

Combined display of all available logs of Hauck LabWiki. You can narrow down the view by selecting a log type, the username (case-sensitive), or the affected page (also case-sensitive).

14:34, 5 June 2025 ESO wikiadmin talk contribs uploaded File:SBA 21.png
14:31, 5 June 2025 ESO wikiadmin talk contribs created page File:SB20.png
14:31, 5 June 2025 ESO wikiadmin talk contribs uploaded File:SB20.png
14:27, 5 June 2025 ESO wikiadmin talk contribs created page Protein expression and purification (Created page with "== '''Expression of HisSUMO-hPOPX2 (SAK) – Part 1/2''' == - '''''Bacteria Sample'':''' #570 BL21 DE3 pRos. pET24a His-SUMO_hPopx2_fulllength - One day prior, aprox. 15:00 : inoculate 50 mL of antibiotics containing media with 3 colonies and incubate o/n at 37°C - 7 mL each of overnight culture were used to inoculate 3 x 333 mL of antibiotics (Kan, cam) containing LB-medium in a 1 L Erlenmeyer flask until OD = 0.7 - When OD is reach...") Tag: Visual edit
14:17, 5 June 2025 ESO wikiadmin talk contribs created page Testexpression in Pichia pastoris (Created page with " == Procedure == * Inoculate 5 mL YPD medium with one '''big''' colony ** in 50 ml falcon tube (secure lid with tape, don’t tighten lid) * Grow o. n. 30 °C, 200 rpm * Centrifuge at 2000 g, 5 min * Resuspend the complete pellet in 5 mL BMMY medium * Measure the OD ** Use a 1:100 dilution for measurement * Start growth ** For starters you don’t need to dilute your suspension ** For optimization purposes you may want to adjust your starting OD *** Recommended is a sta...") Tag: Visual edit
14:07, 5 June 2025 ESO wikiadmin talk contribs created page Transformation of Pichia pastoris (Created page with " == Procedure == * Linearize 20 µg of plasmid of interest with EcoRV * Purify via EtOH precipitation: ** Adjust sample volume to 50 µL (MilliQ) ** Add 5 µL sodium acetate, vortex ** Add 125 µL ice cold EtOH abs.; Incubate on ice for 15 min ** Centrifuge 30 min, 14’000 rpm, 4 °C ** Discard the supernatant ** Wash the pellet with 500 µL colt EtOH 70 % ** Centrifuge 15 min, 14’000 rpm, 4 °C ** Remove the supernatant, dry pellet at RT ** Dissolve in 15-25 µL Mil...") Tag: Visual edit
14:04, 5 June 2025 ESO wikiadmin talk contribs created page Generation of competent Pichia pastoris (Created page with "{{Protokoll-Mix|Inhalt=Wildtype Pichia Pastoris yJC100, grown on YPD agar plate *YPD medium (yeast extract (10 g/L), peptone (20 g/L), glucose (20 g/L)) *Transformation buffer (Tris (10 mM) pH 7,5, Sorbitol (0,6 M), LiAc (100mM), DTT (10 mM)) *Sorbitol (1M) *Inoculation loop *Erlenmeyer flask 500 mL *Spectrometer for OD measurement *50 mL falcons}} ===== Procedure ===== - Inoculate 5 mL of YPD medium with one big colony - Grow 8 h at 30 °C, 200 rpm...") Tag: Visual edit
13:56, 5 June 2025 ESO wikiadmin talk contribs created page Purification of GST-fusion proteins (Created page with "== Procedure == The frozen bacteria pellets are thawed in 5 ml T-buffer on ice and resuspended. Afterwards the samples are incubated with 1 ml lysozyme for 30 min. The cells are broken through the addition of 20 ml lysis buffer and the treatment of ultrasonic sound (impulse 50%, 20sec, 3-5x) on ice water. 300 µl sepharose-solution is added to the suspension and incubated for 10 min with head-over-head rotation. Afterwards the lysate is centrifuged for 20 min at 16000 r...") Tag: Visual edit
13:53, 5 June 2025 ESO wikiadmin talk contribs created page Expression of GST-fusion proteins (Created page with "{{Protokoll-Mix|Inhalt=* *LB-Medium (10g Bacto-Trypton, 5g yeast extract, 5g NaCl, pH 7.0, ad 1l A. bidest) *Ampicillin (100 µg/ml) *IPTG (100 mM Isopropylthiogalactosid) *2xSDS sample buffer 125mM Tris-HCl (pH 6.8), 10% (v/v)-β- *Mercaptoethanol, 5% (w/v) SDS, 0.1% (w/v) Bromphenolblau, 20% (v/v) glycerin *T-buffer 100mM Tris (pH 8.0), 5 mM EDTA *GST-antibody}} == Procedure == === '''Test expression:''' === After cloning, the constructs of the desired GST-fusion pr...") Tag: Visual edit
13:50, 5 June 2025 ESO wikiadmin talk contribs created page Bacterial Pulldown with soluble receptors (Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=PBS (1x) – 24g NaCl, 0.6g KCl, 3.45g Na2HPO4*7H2O, 0.6g KH2PO4, pH 7.4, ad 1l A.bidest Cell culture supernatants (e.g. OptiMEM) harvested from 293cells transiently transfected with constructs encoding soluble CEACAM- ectodomains fused to GFP Polyclonal rabbit anti-GFP antibody (AG Hauck) 2xSDS sample buffer 125mM Tris-HCl (pH 6.8), 10% (v/v) β-Mercaptoethanol, 5% (w/v) SDS, 0.1% (w/v) Bromphenolblau, 20% (v/v) Glycerin}}...") Tag: Visual edit
13:48, 5 June 2025 ESO wikiadmin talk contribs created page Opsonizing bacteria with Fc fusion proteins (Created page with "Bacteria are stained with CFSE and resuspended in PBS. 0.3 bacteria (OD 550) are incubated with either 0.05 mg of purified fusion proteins of 1ml of the unpurified fusion protein for 1 h under rotation at 4°C. As a positive control bacteria are incubated with 500 µl of anti-gonococcal sera (heat-inactivated for 1 h at 56°C) and 500 µl PBS. As a negative control bacteria are incubated with 1 ml of PBS. After the incubation the bacteria are washed and resuspended in 1...") Tag: Visual edit
12:52, 5 June 2025 ESO wikiadmin talk contribs created page File:WB MHE.png
12:52, 5 June 2025 ESO wikiadmin talk contribs uploaded File:WB MHE.png
12:43, 5 June 2025 ESO wikiadmin talk contribs created page Immunoprecipitation (Created page with "This is an original excerpt from MHE's digital labbook from 25 April 2025. {{Protokoll-Box}} = Immunoprecipitation = - THP-1 Cells (WT, PPM1F KO (clone D7), PPM1F KO + PPM1F WT (clone C9), PPM1F KO+ PPM1F D360A (clone A5)) were count and 3x106 cells were lysed using 1000 µl RIPA lysis buffer (10 µg/ml Aprotinin,10 µM Benzamidin, 5 µg/ml Leupepin, 1 µM PMSF, 10 nM pNPP, 2 µM cytochalasinD, cpd26, 1x Phosstop) - Cells were vortexted, s...") Tag: Visual edit
12:28, 5 June 2025 ESO wikiadmin talk contribs created page Bio-Rad GelDoc (Created page with "= Bio-Rad GelDoc XR+ System - Overview = == What is the GelDoc XR+ System? == The GelDoc XR+ System from Bio-Rad is a reliable gel documentation system designed for high-quality imaging of nucleic acid and protein gels. This system combines proven CCD camera technology with versatile illumination options to provide consistent, publication-quality images for routine gel documentation needs. == Main Applications == === Nucleic Acid Gel Documentation === * '''Agarose ge...") Tag: Visual edit
12:24, 5 June 2025 ESO wikiadmin talk contribs created page Bio-Rad ChemiDoc Touch (Created page with "= Bio-Rad ChemiDoc Touch System - Overview = == What is the ChemiDoc Touch System? == The ChemiDoc Touch System from Bio-Rad is a versatile imaging system for documentation and analysis of gels and blots. It combines high-resolution CCD camera technology with intuitive touchscreen operation and enables capture of fluorescence, chemiluminescence, and visible light images. == Main Applications == === Gel Documentation === * '''Agarose gels:''' DNA/RNA electrophoresis w...") Tag: Visual edit
12:17, 5 June 2025 ESO wikiadmin talk contribs created page Thermo Scientific Varioskan Flash eng (Created page with "This page is also available in german 🇩🇪. = Varioskan Flash - Basic Operation Guide = == Overview == The Varioskan Flash is a multimode microplate reader for various detection methods (absorbance, fluorescence, luminescence, time-resolved fluorescence). This guide provides a basic overview - '''the manual remains essential for detailed protocols!''' == Preparation == === 1. Power on the instrument === * Activate main power...") Tag: Visual edit
12:11, 5 June 2025 ESO wikiadmin talk contribs created page Thermo Scientific Varioskan Flash (Created page with "Diese Seite ist auch auf Englisch 🇬🇧 verfügbar. = Varioskan Flash - Grundlegende Bedienung = == Überblick == Der Varioskan Flash ist ein Multimode-Mikroplatten-Reader für verschiedene Detektionsmethoden (Absorbanz, Fluoreszenz, Lumineszenz, Zeit-aufgelöste Fluoreszenz). Diese Anleitung bietet einen ersten Überblick - '''das Handbuch bleibt unverzichtbar für detaillierte Protokolle!''' == Vorbereitung == === 1. Ger...") Tag: Visual edit
11:23, 5 June 2025 ESO wikiadmin talk contribs created page File:Wiki logo.png
11:23, 5 June 2025 ESO wikiadmin talk contribs uploaded File:Wiki logo.png
10:02, 5 June 2025 ESO wikiadmin talk contribs created page File:Lab Biosafety Level 1.png
10:02, 5 June 2025 ESO wikiadmin talk contribs uploaded File:Lab Biosafety Level 1.png
09:46, 5 June 2025 ESO wikiadmin talk contribs uploaded a new version of File:AG Hauck Group photo September 2023.jpg
16:23, 3 June 2025 ESO wikiadmin talk contribs created page File:Cellculture meme.jpg
16:23, 3 June 2025 ESO wikiadmin talk contribs uploaded File:Cellculture meme.jpg
16:22, 3 June 2025 ESO wikiadmin talk contribs created page S1 - Cell culture guidelines (Created page with "'''Getting started:''' * Turn on hood, open bench  needs several minutes, wait for green light * Tie back hair, push back lose sleeves; disinfect hands & arms, put on gloves (can be reused) * Wipe down bench and window with 70% EtOH * Gather all material you need, spray with 70% EtOH and wipe down before placing under hood * Arrange items under hood towards the back of the bench ⇒Keep in mind that you should work a minimum of 15cm inward from the outside edge of th...") Tag: Visual edit
13:52, 3 June 2025 ESO wikiadmin talk contribs created page File:AG Hauck Group photo September 2023.jpg
13:52, 3 June 2025 ESO wikiadmin talk contribs uploaded File:AG Hauck Group photo September 2023.jpg
13:03, 3 June 2025 ESO wikiadmin talk contribs created page Laborsicherheit (Created page with "== '''Schutzkleidung''' == In den Laboren ist bei allen Arbeiten IMMER ein geschlossener Labormantel und eine Schutzbrille zu tragen. Besondere Vorsicht ist bei Arbeiten mit flüssigem Polyacrylamid (Gießen von SDS-PAGE-Gelen) und bei Arbeiten mit Säuren/Laugen walten zu lassen. Zusätzlich zu Labormantel und Schutzbrille sind hierbei Handschuhe zu tragen. == '''Gas''' == Laborgas ist zentral in Raum ML in einem Sicherheitsschrank untergebracht. Die Gasversorgung soll...") Tag: Visual edit
09:49, 3 June 2025 ESO wikiadmin talk contribs created page File:Labetiquette fileserver.jpg
09:49, 3 June 2025 ESO wikiadmin talk contribs uploaded File:Labetiquette fileserver.jpg
09:28, 2 June 2025 ESO wikiadmin talk contribs created page FBA-staining of bacteria (Created page with "{{Protokoll-Mix|Inhalt=(5-(6)-carboxyfluorescein succinimidyl ester (CFSE, FAM SE, Fluorescein) NHS-Biotin PBS++ (PBS, 45 nM CaCl 2, 35 nM MgCl 2) 4% Paraformaldehyde (PFA) Blocking solution (10% FCS in PBS ++) PBS Streptavidin-AlexaFluor647}} == Procedure == * Dilute 0.2 mg NHS-Biotin in 1 ml PBS (prewarm Biotin to RT before opening the tube) * Collect bacteria from plate in PBS, centrifuge for 4 min, 4000 rpm, and resuspend the pellet in 700 l PB...") Tag: Visual edit
09:04, 2 June 2025 ESO wikiadmin talk contribs created page Gentamicin Protection Assay (Created page with "== Procedure == * Aspirate cell culture medium with a pasteur pipette * Rince attaching cells carefully with cold PBS, aspirate PBS * Add 1.5 mL Trypsin/EDTA and study the detachment of the cells in the microscope * Inactivate Trypsin/EDTA with 3.5 mL medium, transfer the cell suspension into a 15 mL centrifuge tube * Pipette 20 mL of the cell suspension into a Neubauer counting chamber * Centrifuge the remaining cells at 600 rpm for 3 min * During centrifugation: Deter...") Tag: Visual edit
08:53, 2 June 2025 ESO wikiadmin talk contribs created page Monitoring bacterial growth (Created page with "{{Protokoll-Mix|Inhalt=Depending on the bacterium you want to analyse, you should determine beforehand: *Growth temperature *Shaking intensity *Generation time *Appropriate medium *Liquid or plate culture}} == Pre-culture == Streak out bacteria one to two days prior to the experiment: two days if you want to do a selection for single clones or if you have to incubate them on antibiotics. Addition of antibiotics usually hampers with the growth - for the determination o...") Tag: Visual edit
08:49, 2 June 2025 ESO wikiadmin talk contribs created page Transformation of Neisseria gonorrhoeae (Created page with "{{Protokoll-Mix|Inhalt=Gonococcal strain which is Opa-postive /Pili positive PPM medium (15g proteose pepton, 5g NaCl, 4g KH 2PO4 1g K2HPO4, 1g soluble starch, ad 1l H20 ph: 7.5) + vitamin mix cloned and sequenced analysed gonococcal DNA which contains the spectinomycin resistance cassette and the Neisserial DNA uptake sequence (DUS) (plasmid DNA}} == Procedure == * Ngo strain (O+/P+) were grown overnight (37°C) on GC agar plates in a humid atmosphere with 5% CO 2 * N...") Tag: Visual edit
08:39, 2 June 2025 ESO wikiadmin talk contribs created page Transformation of plasmids in E. coli (Created page with " == Procedure == * Before transformation, appropriate aliquots of competent bacteria should be thawn on ice. * Add the plasmid DNA to 100 μl competent E. coli in a snap-cap tube and mix well. * Incubate the samples 30 min on ice. * Transfer samples for 75 seconds to a 42°C water bath or heating block and quickly back on ice. * Add 1 ml LB-Medium and incubate the bacteria for 1h at 37°C (220 rpm). * Transfer the bacterial suspension from the snap-cap tube to Eppendorf...") Tag: Visual edit
15:58, 30 May 2025 ESO wikiadmin talk contribs created page MediaWiki:Common.js (Created page with "Any JavaScript here will be loaded for all users on every page load.: // Real Example Tab auf allen Seiten hinzufügen $(document).ready(function() { var realExampleTab = $('<li id="ca-realexample"><a href="/wiki/Real_Example" title="Real Example">Real Example</a></li>'); $('#p-views ul').append(realExampleTab); });")
15:46, 30 May 2025 ESO wikiadmin talk contribs created page InFusion-cloning (Fa. Clontech) (Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=InFusion Kit Becton-Dickinson Aqua bidest Linearised vector (pDNR-Dual), 100ng/1µl (~4900bp) Fragment (10-50ng/µl) -> Subsequent method: Transformation}} == Procedure == * fragment concentration (fragment : pDNR-Dual  2:1 ratio [fmol]) * solution of a dry-down reaction in 102 µl aqua bidest * add 14 µl 10x In-Fusion-buffer and 2 µl pDNR-Dual (200 ng) * disperse to 15 tubes (8 µl per tube) and add 0.5 -1 µl of...") Tag: Visual edit
15:43, 30 May 2025 ESO wikiadmin talk contribs created page Cre-Lox recombination of plasmids (Created page with "== Procedure == * Dilute Cre-recombinase [10 U/µl] with 1x Cre-recombinase-buffer 1:10 [1 U/µl] * Recombination reaction contains: ** 1 µl 10x Cre-recombination-buffer 6µl aqua bidest. ** 1 µl (100ng) donor vector (e.g. pDNR-dual gene X) ** 1 µl (200ng) acceptor vector (e.g. pLPS3’-EGFP) ** 2 µl Cre-recombinase * Incubation for 45 min at 37° C, meanwhile starting heating-unit * Heat inactivation for 10 min at 70° C and subsequent cooling to room temperature...") Tag: Visual edit
15:40, 30 May 2025 ESO wikiadmin talk contribs created page Site directed mutagenesis (Created page with "== Procedure == * Prepare the PCR reaction mix according to the hot start procedure. Include one negative control sample without Pfu polymerase (incubate sample at the lowest annealing temperature of your gradient) * Perform gradient PCR using the following thermal cycling conditions: {| class="wikitable" !Step !Temperature °C !Time !No. of cycles |- |Initial denauturation |94 |1-3 min |1 |- |Hot Start / add enzyme |94 |pause |1 |- |Denauturation |94 |20 sec | rowspan...") Tag: Visual edit
15:33, 30 May 2025 ESO wikiadmin talk contribs created page Cloning of guideRNA-Oligo into pLentiCRISPRv2 (Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=gRNA Oligos (sense/antisense; designed using ChopChop) Vector pBluescript_U6-MCS digested with BbsI (Plasmid #3468) T4 DNA ligase; 10x Ligase buffer E. coli NovaBlue}}")
15:30, 30 May 2025 ESO wikiadmin talk contribs created page Cloning of guideRNA-Oligo into pBluescript U6-MCS (Created page with "== Procedure == '''gRNA Oligo annealing''' * annealing reaction contains: 1 µl sense primer (stock solution 100 µM), 1 µl antisense primer (stock solution 100 µM), 5 µl NEB buffer 2 and 43 µl ddH2O * Annealing reaction occurs in thermocycler with the corresponding program: {| class="wikitable" |95°C |4 minutes |- | colspan="2" |Start loop, 54x, 94°C, 1 min (-1°C/loop) |- |8°C |Store forever |} == Preparation of BbsI digested vector pBluescript_U6...") Tag: Visual edit
15:21, 30 May 2025 ESO wikiadmin talk contribs created page File:Protokoll guideRNA.jpg
15:21, 30 May 2025 ESO wikiadmin talk contribs uploaded File:Protokoll guideRNA.jpg
15:18, 30 May 2025 ESO wikiadmin talk contribs created page Design of guideRNA-Oligos (gRNA) for CRISPR/Cas gen. editing (Created page with "{{Protokoll-Mix|Inhalt= 20 mM dNTPs 10x Taq/Pfu buffer 2 Primers 10 μM DNA template, 100 – 200 ng/μL DNA polymerases (Pfu, Taq)}}") Tag: Visual edit: Switched
12:13, 30 May 2025 ESO wikiadmin talk contribs created page Cloning of shRNA into lentiviral vector pLKO.1 (Created page with "== Procedure == === Primer annealing === * annealing reaction contains: 1 µl sense primer (stock solution 100 µM), 1 µl antisense primer (stock solution 100 µM), 5 µl NEB buffer 2 and 43 µl ddH2O * Annealing reaction occurs in thermocycler with the corresponding program:") Tag: Visual edit
12:04, 30 May 2025 ESO wikiadmin talk contribs created page LIC Cloning (Ligation independent cloning) (Created page with " == Procedure == Design primers compatible for cloning in vector containing the LIC sequence (e.g. pDNR Dual LIC) and perform PCR. Separate 4 µl of PCR reaction via agarose gel electrophoresis to check PCR product. Many unspecific bands besides the desired one require gel separation of the PCR sample and extraction of the desired band. If necessary, 2-3 PCR reactions can be pooled to gain more material. Add 10x NEB2 Buffer to the PCR sample to reach 1x end concentration...") Tag: Visual edit: Switched
11:48, 30 May 2025 ESO wikiadmin talk contribs created page File:LIC sequences.jpg (Fig. 1: Sequences to include in forward and reverse primers intended for LIC cloning. Codons representing the proper reading frame after transfer from pDNR-dual LIC are indicated. NNN indicates the start of the sequence complementary to your target sequence. Please remem- ber to use the antisense sequence in the reverse primer.)
11:48, 30 May 2025 ESO wikiadmin talk contribs uploaded File:LIC sequences.jpg (Fig. 1: Sequences to include in forward and reverse primers intended for LIC cloning. Codons representing the proper reading frame after transfer from pDNR-dual LIC are indicated. NNN indicates the start of the sequence complementary to your target sequence. Please remem- ber to use the antisense sequence in the reverse primer.)
11:29, 30 May 2025 ESO wikiadmin talk contribs created page Template:Abbildung (Created page with "<div style="text-align: center; margin: 25px 0; padding: 15px; border: 1px solid #a2a9b1; border-radius: 4px; background: #f8f9fa;"> [[Datei:{{{datei}}}|{{{groesse|400px}}}]] <div style="margin-top: 10px; font-size: 13px; color: #555; line-height: 1.4;"> {{{titel|Fig. 1:}}} {{{beschreibung|Bildbeschreibung}}} </div> </div>")

Main public logs

Navigation menu

Search