User contributions for ESO wikiadmin

30 May 2025

• 12:1312:13, 30 May 2025 diff hist +290 N 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
• 12:1212:12, 30 May 2025 diff hist +478 LIC Cloning (Ligation independent cloning) No edit summary current Tag: Visual edit
• 12:1112:11, 30 May 2025 diff hist +345 LIC Cloning (Ligation independent cloning) No edit summary Tag: Visual edit
• 12:0412:04, 30 May 2025 diff hist +1,516 N 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
• 11:5611:56, 30 May 2025 diff hist +100 Designing PCR-Primers for LIC Cloning (Ligation independent cloning) No edit summary current Tag: Visual edit
• 11:5611:56, 30 May 2025 diff hist +1,131 Designing PCR-Primers for LIC Cloning (Ligation independent cloning) No edit summary Tag: Visual edit
• 11:5511:55, 30 May 2025 diff hist +147 Designing PCR-Primers for LIC Cloning (Ligation independent cloning) No edit summary
• 11:5411:54, 30 May 2025 diff hist −1 Designing PCR-Primers for LIC Cloning (Ligation independent cloning) No edit summary
• 11:5111:51, 30 May 2025 diff hist +666 Designing PCR-Primers for LIC Cloning (Ligation independent cloning) No edit summary
• 11:4811:48, 30 May 2025 diff hist +343 N 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. current
• 11:2911:29, 30 May 2025 diff hist +341 N 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;"> <strong>{{{titel|Fig. 1:}}}</strong> {{{beschreibung|Bildbeschreibung}}} </div> </div>" current
• 11:2811:28, 30 May 2025 diff hist +652 N Designing PCR-Primers for LIC Cloning (Ligation independent cloning) Created page with " == Procedure == Delineate the coding sequence (full-length or partial sequence), which you want to amplify. Sense and antisense primers should have around 18 to 22 complementary bases, depending on the AT-content (there should be 9-10 Gs or Cs in the primer- cDNA complementary region). Both primers should ideally end with a 3’-GC clamp (two G or C bases). In the antisense primer, add a STOP codon, if you want to create a protein with an N- terminal tag; omit the STOP..." Tag: Visual edit
• 11:2511:25, 30 May 2025 diff hist +1,149 Isolation of DNA from agarose gel No edit summary current Tag: Visual edit
• 11:2311:23, 30 May 2025 diff hist +146 N Isolation of DNA from agarose gel Created page with "{{Protokoll-Mix|Inhalt= 20 mM dNTPs<br>10x Taq/Pfu buffer<br>2 Primers 10 μM<br> DNA template, 100 – 200 ng/μL<br>DNA polymerases (Pfu, Taq)}}" Tag: Visual edit: Switched
• 11:2211:22, 30 May 2025 diff hist +583 PCR purification No edit summary current Tag: Visual edit
• 11:2111:21, 30 May 2025 diff hist +122 N PCR purification Created page with "{{Protokoll-Mix|Inhalt=TAE buffer (1x) 4.84 g Tris-Base, 1 mM EDTA, 1.14 mL acetic acid, fill 1 L with A.bidest. Agarose}}" Tag: Visual edit: Switched
• 11:2011:20, 30 May 2025 diff hist +108 Setting up quantitative real-time PCR qRT-PCR No edit summary current Tag: Visual edit
• 11:1711:17, 30 May 2025 diff hist +436 Setting up quantitative real-time PCR qRT-PCR No edit summary
• 11:1611:16, 30 May 2025 diff hist +505 Setting up quantitative real-time PCR qRT-PCR No edit summary Tag: Visual edit
• 10:0110:01, 30 May 2025 diff hist +38 Setting up quantitative real-time PCR qRT-PCR No edit summary Tag: Visual edit: Switched
• 10:0010:00, 30 May 2025 diff hist +28 N Setting up quantitative real-time PCR qRT-PCR Created page with "{{Pipetten-Checklist-8step}}" Tag: Visual edit
• 09:5909:59, 30 May 2025 diff hist +3,606 N Template:Pipetten-Checklist-8step Created page with "<div style="display: inline-block; border: 1px solid #a2a9b1; border-radius: 2px; margin: 10px 0; max-width: fit-content; background: #ffffff; box-shadow: 0 1px 3px rgba(0,0,0,0.1);"> <div style="background: #eaecf0; padding: 8px 12px; border-bottom: 1px solid #a2a9b1; font-weight: bold; color: #333; font-size: 14px;"> <div>✓ {{{titel|Pipettierschema - Checklist}}}</div> </div> <div style="padding: 15px; color: #333; font-size: 13px;"> <div> <table..." current
• 09:5409:54, 30 May 2025 diff hist +336 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) →Touch-down PCR: current Tag: Visual edit
• 09:5109:51, 30 May 2025 diff hist −478 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary
• 09:5109:51, 30 May 2025 diff hist −257 PCR approach in 20 μL with hot-start (qualitative PCR) No edit summary current Tag: Visual edit
• 09:4609:46, 30 May 2025 diff hist +1,806 PCR approach in 20 μL with hot-start (qualitative PCR) No edit summary
• 09:4109:41, 30 May 2025 diff hist +64 N PCR approach in 20 μL with hot-start (qualitative PCR) Created page with "{{Protokoll-Mix|Inhalt= The same as for PCR approach in 50 µL}}"
• 09:3909:39, 30 May 2025 diff hist 0 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 09:3809:38, 30 May 2025 diff hist −30 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 09:3709:37, 30 May 2025 diff hist +78 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 09:3609:36, 30 May 2025 diff hist +850 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 09:3309:33, 30 May 2025 diff hist +4,227 N Template:Pipetten-Checklist-flexibel Created page with "<div style="display: inline-block; border: 1px solid #a2a9b1; border-radius: 2px; margin: 10px 0; max-width: fit-content; background: #ffffff; box-shadow: 0 1px 3px rgba(0,0,0,0.1);"> <div style="background: #eaecf0; padding: 8px 12px; border-bottom: 1px solid #a2a9b1; font-weight: bold; color: #333; font-size: 14px;"> <div>✓ {{{titel|Pipettierschema - Checklist}}}</div> </div> <div style="padding: 15px; color: #333; font-size: 13px;"> <div> <table..." current
• 09:3209:32, 30 May 2025 diff hist +238 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary
• 09:2809:28, 30 May 2025 diff hist −4 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 09:2809:28, 30 May 2025 diff hist +30 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary
• 09:2709:27, 30 May 2025 diff hist −620 Template:Pipetten-Checklist No edit summary current
• 09:2409:24, 30 May 2025 diff hist +114 Template:Pipetten-Checklist No edit summary
• 09:2109:21, 30 May 2025 diff hist −1 AG Hauck - LabWiki No edit summary

29 May 2025

• 18:2418:24, 29 May 2025 diff hist +229 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 18:2118:21, 29 May 2025 diff hist +44 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary Tag: Visual edit
• 18:2118:21, 29 May 2025 diff hist +253 Template:Pipetten-Checklist No edit summary
• 18:1818:18, 29 May 2025 diff hist +3,239 N Template:Pipetten-Checklist Pipettierschmema allgemein
• 18:1618:16, 29 May 2025 diff hist +3,395 N Template:Pipettierschema Created page with "<div style="display: inline-block; border: 1px solid #a2a9b1; border-radius: 2px; margin: 10px 0; max-width: fit-content; background: #ffffff; box-shadow: 0 1px 3px rgba(0,0,0,0.1);"> <div style="background: #eaecf0; padding: 8px 12px; border-bottom: 1px solid #a2a9b1; font-weight: bold; color: #333; font-size: 14px;"> <div>✓ {{{titel|Pipettierschema - Checklist}}}</div> </div> <div style="padding: 15px; color: #333; font-size: 13px;"> <div> <table..." current
• 18:1318:13, 29 May 2025 diff hist +11 PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) No edit summary
• 18:1218:12, 29 May 2025 diff hist +497 N PCR approach in 50 μL with hot-start and/or touch-down (preparative PCR) 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)}} == Procedure == As usually more than one PCR sample is run simultaneously, reaction and enzyme mix are prepared separately. The enzyme mix is added to the reaction mix after starting the PCR and after pre-heating the reaction mix to 94 ºC (hot start). A typical 50 μL approach that yields enough material for later cloning procedures..." Tag: Visual edit
• 18:0418:04, 29 May 2025 diff hist +2,490 N Design principles for PCR primers used for amplification and cloning of cDNA Created page with "'''Restriction Enzyme Sites:''' For conventional cloning with restriction enzymes, the recognition sites are added to the 5’ end of the primer. Check that the involved '''restriction enzymes do NOT cut within the cloned cDNA''' and that they do '''NOT cut the vector outside the intended position'''. Make sure to preserve the reading frame, if the DNA is cloned 3’ or 5’ of existing ORFs (e.g. GFP) in the vector. Add 3 to 4 meaningless nucleotides (e.g. ATAA-) at the..." current Tag: Visual edit
• 18:0218:02, 29 May 2025 diff hist −245 Agarose gel electrophoresis No edit summary current Tag: Visual edit
• 18:0118:01, 29 May 2025 diff hist +105 Agarose gel electrophoresis No edit summary Tag: Visual edit
• 18:0018:00, 29 May 2025 diff hist +19 Template:Protokoll-Mix No edit summary current
• 17:5917:59, 29 May 2025 diff hist +18 Agarose gel electrophoresis No edit summary Tag: Visual edit