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- 17:06, 25 September 2019 Neonatal mouse cardiomyocyte culture (hist) [715 bytes] Stephan Lange (Talk | contribs) (Created page with "== Isolation & culture of neonatal mouse cardiomyocytes == The method has been published in [https://www.ncbi.nlm.nih.gov/pubmed/24056408 JoVE (PMID: 24056408)]. File:NMC...")
- 16:10, 31 July 2018 Pierce BCA Protein Concentration (hist) [1,865 bytes] Mclark (Talk | contribs) (Created page with "==Summary== We use this method measure the amount of protein we have in our unknown samples ===Method=== # *Place samples in order on ice* # Use the sonicator to break up all...")
- 00:59, 31 July 2018 Transfecting C2C12 Cells (hist) [1,714 bytes] Mclark (Talk | contribs) (Created page with "=== Transfecting C2C12 Cells=== * siRNA against Cullin 3 * siRNA with other sequence ===Materials=== * Two tubes (1.7ml) per well (12 tubes total) * 6 for control * 6 for tr...")
- 20:12, 23 July 2018 Measuring RNA Concentration (hist) [1,153 bytes] Mclark (Talk | contribs) (Created page with "===Summary=== In order to quantify the concentration of RNA we have in our sample we use a Nanodrop * Nanodrop is located in the hallway * Need kimwipes and milli Q water (M...")
- 15:43, 23 July 2018 Extraction of RNA (hist) [3,235 bytes] Mclark (Talk | contribs) (Created page with "===Extracting RNA post stimulation and collection=== '''EXAMPLE:''' Starting w/ four samples (before, 6hr, 12hrs, 24hrs) # Use the hood in the hallway, remove any stuff that...")
- 15:41, 23 July 2018 Creating a Lower Buffer Solution (hist) [863 bytes] Mclark (Talk | contribs) (Created page with "===Making 1.5M Lower Buffer Solution=== *500 total to make # 91g of tris base (into a beaker with a stir bar) # 350 ml of milli Q (from the tap not reservoir) measure in a gr...")
- 21:23, 20 July 2018 Transfecting Cells (hist) [1,523 bytes] Mclark (Talk | contribs) (Created page with "==Summary== Cos 1 (African green monkey, from kidney cells). Cells are heterozygous, they all look different. We are transfecting Cos 1, 6 cm plates * Make sure everything is...")
- 21:16, 20 July 2018 Designing Oligonucleotides for qPCR (hist) [2,460 bytes] Mclark (Talk | contribs) (Created page with "==Summary== We design primers get an optimal PCR. It is an important step. If you use any primers without design, the amplification may not work or give you skewed results =...")
- 21:04, 20 July 2018 Seeding C2C12 Cells (hist) [2,293 bytes] Mclark (Talk | contribs) (Created page with "==Seeding C2C12 cells== Check how confluent cell dishes are. Mark the one that will be used for seeding (if there is more than one dish) choose the medium confluent dish and t...")
- 20:18, 19 July 2018 Collection of Proteins and RNA Post- Stimulation (hist) [1,451 bytes] Mclark (Talk | contribs) (Created page with "===Collecting proteins at each stimulation time point=== # Warm solutions needed (PBS, lysis buffer) # Clean cell scrapers in DI water, place in hood # Collect ice for the sam...")
- 20:04, 19 July 2018 Stimulation of C2C12 Cells with Neural Agrin (hist) [1,315 bytes] Mclark (Talk | contribs) (Created page with "==Stimulation of C2C12 cells with Agrin== As mentioned before: -PBS is the control (found in the freezer, bottom shelf, agrin box, PBS 0.1% BSA 1000x) -Neural agrin (x1000) is...")
- 20:01, 19 July 2018 Scanning PCR Gel (hist) [635 bytes] Mclark (Talk | contribs) (Created page with "==Scanning PCR gel== # After the gel is done running, disconnect the power source and remove gel onto a piece of paper or a paper towel # Dry the gel a little and place into t...")
- 22:57, 18 July 2018 Genotyping PCR (hist) [2,453 bytes] Mclark (Talk | contribs) (Created page with "==Summary== DNA is amplified by three steps: denaturation, annealing, and elongation. The wild type control (WT) is negative for both CRE and Cullin 3 CRE positive and cullin...")
- 22:52, 18 July 2018 Making 2% Agarose Gel for PCR Analysis (hist) [863 bytes] Mclark (Talk | contribs) (Created page with "==Making 2% Agarose Gel for PCR Analysis== ===Materials=== * Agarose powder (kept in cabinet) * 1x TAE liquid (by sink) ===Methods=== # Start by taping the gel mold (found b...")
- 22:50, 18 July 2018 Making 4% Agarose Gel for PCR Analysis (hist) [864 bytes] Mclark (Talk | contribs) (Created page with "==Making 4% Agarose Gel for PCR Analysis== ===Materials=== * Agarose powder (kept in cabinet) * 1x TAE liquid (by sink) ===Methods=== # Start by taping the gel mold (found b...")
- 22:49, 18 July 2018 Making 2% Agarose Gel for PCR analysis (hist) [906 bytes] Mclark (Talk | contribs) (Created page with "==Making 2% Agarose Gel for PCR Analysis== ===Materials=== * Agarose powder (kept in cabinet) * 1x TAE liquid (by sink) ===Methods=== # Start by taping the gel mold (found b...")
- 22:46, 18 July 2018 Freezing C2C12 and Satellite Cells (hist) [1,833 bytes] Mclark (Talk | contribs) (Created page with "==Freezing C2C12 and Satellite cells== ===Methods=== * Mr. Freeze box with isopentane * Cryotube (in tissue culture box) * Warm PBS, basic FGF *Trypsin * Gelatin coated dish...")
- 22:39, 18 July 2018 Creating Cell Culture Media (hist) [1,260 bytes] Mclark (Talk | contribs) (Created page with "==How to Make Cell Culture Media for C2C12 and Satellite Cells== ===Differentiation Media for C2C12 cells=== * 200 ml DMEM * 4 ml horse serum (2%) * 3 ml penicillin/ streptomy...")
- 22:30, 18 July 2018 Passaging of C2C12 cells (hist) [900 bytes] Mclark (Talk | contribs) (Created page with "==Making a New Passage C2C12 cells== ===Summary=== We passage cells to keep them alive and growing in the incubator in the cell culture room. Generally cells should be passage...")
- 22:16, 18 July 2018 Changing Proliferation Media to Differentiation Media (hist) [641 bytes] Mclark (Talk | contribs) (Created page with "==C2C12 - Changing Proliferation Media to Differentiation Media== ===Summary=== In order to continue myotube formation 'in vitro' we must change the proliferation medium (afte...")
- 22:11, 18 July 2018 Developing Your Immunoblot Membrane (hist) [1,959 bytes] Mclark (Talk | contribs) (Created page with "==Developing your Immunoblot Membrane== ===Materials=== *A flash drive *A tube rack *Sheet protectors *PICO developing solution *15 ml tube *Ethanol + KimWipes towels # Afte...")
- 21:54, 18 July 2018 Preparing Secondary Antibodies for Immunoblots (hist) [2,007 bytes] Mclark (Talk | contribs) (Created page with "==Preparing Secondary Antibodies for Immunoblots== ===Summary=== As mentioned the secondary antibody is used to visualize what you are looking for, it is an amplificator. We a...")
- 21:20, 18 July 2018 Preparing Primary Antibodies for Immunoblots (hist) [2,187 bytes] Mclark (Talk | contribs) (Created page with "==Preparing Primary Antibodies for Immunoblot== ===Summary=== Make sure the proteins you are identifying are not the same molecular weight. Antibodies are important because...")
- 21:16, 18 July 2018 Ponceau Staining of Protein Blots (hist) [2,364 bytes] Mclark (Talk | contribs) (Created page with "==Ponceau Staining of Protein Blots== ===Summary=== In order to ensure the transfer went through you will stain your membrane in ponceau solution. ===Materials=== * Pre-made...")
- 21:12, 18 July 2018 Ponceau staining of protein blots (hist) [2,388 bytes] Mclark (Talk | contribs) (Created page with "==Ponceau Staining of Protein Blots== ===Summary=== In order to ensure the transfer went through you will stain your membrane in ponceau solution. ===Materials=== * Pre-made...")
- 21:01, 18 July 2018 Loading a Protein Gel (hist) [1,423 bytes] Mclark (Talk | contribs) (Created page with "(Identify how much protein you will have to load in the wells (excel sheet)) # Take the gel apparatus and place it onto the running adapter (positive and negative screws) make...")
- 21:00, 18 July 2018 Running a Protein Gel (hist) [324 bytes] Mclark (Talk | contribs) (Created page with "# Connect (green) power top onto the running adapter. Ensure the screws are on tight and top matches with screws: black to black, red to red. # Set power supply to 100 volts...")
- 20:57, 18 July 2018 Transferring a Protein Gel (hist) [1,373 bytes] Mclark (Talk | contribs) (Created page with "# Once run is complete (blue is running out of gel) switch off power supply # Place transfer buffer into a container (found on drying rack) '' Transfer buffer: 28.8g glycine,...")
- 20:53, 18 July 2018 Transfering a Protein Gel (hist) [0 bytes] Mclark (Talk | contribs) (Created page with "==Transferring a Protein Gel== # Once run is complete (blue is running out of gel) switch off power supply # Place transfer buffer into a container (found on drying rack) ''...")
- 20:48, 17 July 2018 Cell Biology (hist) [593 bytes] Mclark (Talk | contribs) (Created page with "== Methods in Cell Biology == === Working with C2C12 cells === * Passaging of C2C12 cells * Changing Proliferation Media to Differentiation Media")
- 20:43, 17 July 2018 SDS-page gels (hist) [7,387 bytes] Mclark (Talk | contribs) (Created page with "== Creating a Protein Gel == === Materials === * Clean casting stand w/ dual gel apparatus * Glass plate holders (w/ white or black knobs) * Additional spacers (1.5) with (...")
- 18:01, 17 July 2018 Libraries (hist) [279 bytes] Stephan Lange (Talk | contribs) (Created page with "== Available small molecule libraries and drug screening cores and resources at the University of California == [http://cddi.ucsd.edu/resources/index.html?_ga=2.80696151.175...")
