https://padlet.com/jjordan248/ve04gv7i75s2ilzh Lab Tests Study Tool - White: Membrane Transport Green: Lab Tests en-us 2023-03-02 16:11:08 UTC 2026-01-19 22:47:36 UTC hello@padlet.com https://padlet.net/icons/png/1f97c.png jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501367682 - Maintains homeostasis by controlling what goes in and what goes out 
- Phospholipid bilayer
   - Heads of lipids: Hydrophilic; polar
- Tails of lipids: Hydrophobic; nonpolar]]> 2023-03-02 16:20:50 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501367682 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501372361 - Molecules move from high to low concentration 
   - Moves with the concentration gradient
- Simple Diffusion
   - Very small non polar molecules can pass through the phospholipid with no help or energy
   - Molecules move from high to low concentration
   - Moves with the concentration gradient
- Facilitated Diffusion 
   - Uses a transport protein because molecule may be too big or polar
   - Does not require energy
- May require a stimulus(usually a ion)]]> 2023-03-02 16:24:06 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501372361 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501377907 -Movement from low to high concentration 
   - Moves against the gradient 
-Requires Energy
   -ATP
- ATP energizes the protein channel to transport molecule against the concentration gradient
- Endocytosis (3 types)
- A type of active transport where the substance fuses with the cell membrane and brings the substance inside the cell in a vesicle

]]> 2023-03-02 16:27:38 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501377907 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501380136 1.) Phagocytosis 
   - Used by amoebas to engulf a substance
2.) Receptor Endocytosis
   - Special receptor proteins capture a specific target molecule
3.) Pinocytosis
- Intake of liquid ]]> 2023-03-02 16:29:18 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501380136 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501381831 - Exocytosis is another form of active transport 
   - Cell expels/releases/removes material out of the cell
- This can include releasing neurotransmitters or proteins]]> 2023-03-02 16:30:29 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501381831 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501388301 - Separates DNA by size 
   - Shorter DNA travels further through the gel
- Utilizes Agarose 
   - Agarose is very porous 
- Needs a buffer
   - This allows the electric current to travel through the gel
- Uses 
   - Crime Scenes 
   - Paternal/Maternal Testing
   - Comparing Species
   - Identifying/Observing Certain Diseases
- When separating DNA, it all has the same charge
   - DNA will move prom negative to positive ends
- DNA Ladder
   - Known DNA sizes that are used as a standard to compare other samples

]]> 2023-03-02 16:35:02 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501388301 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501391180 - Each line on the gel is a band of DNA
   - Thicker bands mean there was a lot of DNA in that specific band 
- DNA samples that have similar banding patterns will be more closely related
   - Closely related organisms have similar DNA, therefore their banding patterns will be similar]]> 2023-03-02 16:37:05 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501391180 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501393825 - Polymerase Chain Reaction 
- Used to make many copies of DNA region to study
- 3 Main Steps
   - Denaturation
   - Annealing
   - Extension

]]> 2023-03-02 16:38:50 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501393825 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501400705 Step 1:
- Denaturation 
   - Heat the strands of DNA to break(denature) them apart into two separate strands
   - This gives room to replicate the desired gene
Step 2:
- Annealing 
   - Cool DNA so primers can bind 
   - Primers bind to areas of the DNA that we want to amplify
   - Two primers needed for each side of DNA
Step 3:
- Extension
   - Heat up DNA to use taq polymerase 
   - Taq polymerase adds nucleotides to extend the primers and replicate DNA
   - This enzyme is adapted to work in high temps
20x-30x:
- These 3 steps are repeated many, many times
- DNA made from a previous cycle is used as a template
   - This ensures that the desired DNA is the only thing getting replicated

]]> 2023-03-02 16:43:25 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501400705 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501403692 - PCR lets us take a small section of DNA and make hundreds of copies
- This small section can be amplified and examined through many processes 
   - For example, gel electrophoresis 
- Commonly used in forensics]]> 2023-03-02 16:45:25 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501403692 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501414425 - Clustered Regularly Interspaced Short Palindromic Repeats
   - First discovered in E. coli and also found in archaea
   - CRISPRs serve as a part of bacterial immune system
- It refers to the various systems that can be programmed to target specific stretches of genetic code
   - This is so it can edit DNA in precise locations
- An enzyme called Cas9(produced by the CRISPR system) will bind to the matching DNA using a "spacer"
   - Spacer: Short RNA sequences that guide the CRISPR system to target DNA
- Cas9 will cut the DNA, shutting the targeted gene off
   - Research suggests CRISPR-Cas9 can be used to target and modify "typos" (aka mutations) in the human genome
   - Ex: Cpf1
PROS
- Scientists can Accelerate Research
- Rapid Diagnostic 
- Correct Mutations
CONS
- Ethics(how far is too far?)
   - Potentially wiping out species, creating bacterial resistance 
- Mutations at cut site or other places in genome]]> 2023-03-02 16:52:21 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501414425 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501417589 - Enzyme Linked Immunosorbent Assay
- Measures antibodies, antigens, proteins, and glycoproteins in a biological sample
- Diagnosis, pregnancy tests, test for contamination, allergens, and disease progression]]> 2023-03-02 16:54:23 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501417589 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501422305 Step 1: Antibody Coating
- Specific capture antibody is immobilized on high protein binding plates
Step 2: Protein Capture
- Samples and standard dilutions are added to wells and will be captured by bound antibodies
Step 3: Detection Antibody
- Specific detection antibody is added to enable detection of captured protein 
   - If protein is absent, this antibody will attach to nothing
Step 4: Detection Conjugate(enzyme)
- Binds to detection antibody (like a signal)
Step 5: Substrate Added
- Changes color to analyze results ]]> 2023-03-02 16:57:35 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501422305 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501427095 Diluting Samples:
- Why diluting the samples is necessary when using the ELISA Test!
   - Can protect against false positives
   - Can also show disease progression 
Interpreting Results:
- Look for color change 
- Amount of color change can be used to signify rate of disease progression 
- More marker detected = More color change]]> 2023-03-02 17:00:45 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501427095 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501506085 - Also known at lmmunoblot
- It’s a form of gel electrophoresis to detect specific proteins
   - Detect protein of interest 
   - Size of proteins 
   - Amount of protein expressed 
- We’re using a charged solution to draw proteins through a gel 
- Proteins are sorted by mass and ability to bind to antibodies
Western Blot Antibodies:
- Antibodies are Y-shaped proteins that bind to a specific target (antigen)
- Western Blotting uses antigens to detect certain proteins out of a large group


]]> 2023-03-02 17:57:50 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501506085 jjordan248 https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501506475 - Proteins in the sample are coated with a detergent to denature(unfold) them and give them a negative charge 
   - We need linear proteins to move through gel
- Preform gel electrophoresis 
   - Once complete, move proteins onto a membrane
- Take completed gel and membrane and sandwich them between filter paper
-Negatively charged proteins will be drawn to the positive end 
   - The membrane is between positive end of chamber and the gel, so the proteins will be drawn onto the membrane
   - They’ll be “blotted” onto the membrane
   - This membrane is an identical copy of the gel
- Why the membrane?
   - Gel is too fragile for next steps 
   - Antibodies could become stuck in the gel and give false results 
- Once proteins have moved onto the membrane, we add antibodies to locate proteins we want to see
   - The antibodies act like a flag on the target protein 
- Once the membrane copies the gel, it’s moved to a chamber and exposed to primary antibodies
   - This primary antibody specifically attaches to the protein we’re looking for
   - Once attached, the membrane is washed so extra antibodies don’t  stick around
- A secondary antibody is then added to the membrane 
   - This secondary antibody attaches to the primary antibody, not the protein
   - Contains a reporter enzyme that will produce light or color
- The reporter enzyme allows us to locate the proteins 
- By adding the antibodies, we only see the proteins we’re looking for on thr funal membrane

]]> 2023-03-02 17:58:07 UTC https://padlet.com/jjordan248/ve04gv7i75s2ilzh/wish/2501506475