All Protocols















ADN_Cas9_Feldan.jpg

CRISPR/CAS9




Coming+Soon.jpg

ELISA



Complete List of Protocols:

  1. The Sterile Workspace

  2. Host Selection Criteria

  3. Culturing Conditions

  4. Mixing Media and Pouring Plates

    1. LB Liquid Media

    2. LB-Agar Media

    3. Antibiotic Stock Solution Mixing

  5. Sterile Technique

  6. DIY Buffer Mixing

    1. Water

    2. Tris-Cl Buffer

    3. Miniprep Buffer Mixing

      1. Buffer P1 (Resuspension Buffer)

      2. Buffer P2 (Lysis Buffer)

      3. Buffer N3 (Neutralisation Buffer)

      4. Buffer PB (Wash Buffer 1)

      5. Buffer PE (Wash Buffer 2)

      6. Elution Buffers (TE, EB, Water)

    4. Agarose Electrophoresis Buffers

      1. TAE Buffer

      2. LAB Buffer

      3. TBE Buffer

    5. Buffer QG (Gel Purification Buffer)

    6. Protein Purification Buffers

      1. Protein Wash Buffer

      2. Protein Elution Buffer

      3. Protein Storage Buffer

    7. SDS-PAGE Buffers

      1. SDS-PAGE 4x Loading Buffer

      2. SDS-PAGE 10x Running Buffer

      3. Coomassie Blue Staining Solution

      4. Destaining Solution

  7. DNA101

  8. Purification of Genomic DNA

    1. Boil Purification of Genomic DNA

    2. Genomic DNA Extraction with CTAB

  9. Plasmids101 - Guide coming soon!

  10. Purification of Plasmid DNA (Miniprep aka. Ethanol Lysis)

  11. Plasmid Insertion

    1. Preparation of Chemically Competent E.coli cells (Rubidium Chloride) for Heat Shock Transformation

    2. Preparation of Chemically Competent E.coli cells (Calcium Chloride) for Heat Shock Transformation

    3. Heat Shock Transformation of Chemically Competent E. Coli

    4. Preparation of Electrocompetent cells of Pseudomonas or E. coli for Electroporation

    5. Electroporation of Electrocompetent cells of Pseudomonas or E. coli for Electroporation

    6. DIY Gene Gun

  12. Agarose Gel Electrophoresis

    1. Buffer QG Gel Purification

    2. Cotton Wool Gel Purification

  13. Gel Illumination & Photography

  14. Restriction Enzyme Digest of Plasmid & Insert DNA, Optional: Dephosphorylation + Purification

    1. Heat Inactivation of Restriction Enzymes

    2. Column Purification of Restriction Enzyme Digest

  15. Ligation & Troubleshooting

    1. Kinase Treatment

  16. Polymerase Chain Reaction (PCR)

  17. Primer Design

  18. Gradient PCR

  19. Confirmation of Edits

    1. Patch Plate and Colony PCR (Junction & Spanning PCR)

    2. Restriction Digest Analysis

    3. Preparation for External Sequencing

    4. In-House Sequencing (Oxford Nanopore)

    5. Interpreting Sequencing Results

    6. Preparation of Glycerol Stocks

  20. Flanking Sequence Addition

  21. Gibson Assembly

  22. Golden Gate Assembly

  23. Site Directed Mutagenesis

  24. GBlock Design

  25. Backbone Selection

  26. CRISPR/CAS9

  27. Proteins101

  28. Induction of Protein Expression

  29. Cell Lysis

    1. Bead-Beating Lysis

    2. Chloroform Lysis of Periplasm

    3. Chemical Lysis

  30. Affinity Tag Purification

    1. His-Tag Protein Purification

      1. Centrifuge & Filter His-Tag Purification

      2. Column Chromatography His-Tag Purification

    2. Car9 Protein Purification

  31. SDS-PAGE

  32. Antibody Production

  33. Aptamer Production

  34. Cardinal Blots

    1. Northern Blot

    2. Eastern Blot

    3. Southern Blot

    4. Western Blot

  35. ELISA

  36. Mass Spec Prep

  37. Cell Assays

  38. RNA101

  39. RNA Purification

  40. RT-PCR

  41. RT-qPCR

  42. In Silica