Polymerase Chain Reaction (PCR)
PCR Is a common lab technique used to make many copies of a DNA sample to work with or store in a laboratory. To learn more about it, check out these links!
https://www.youtube.com/watch?v=iQsu3Kz9NYo
https://media.hhmi.org/biointeractive/vlabs/bacterial_id/index.html (interactive PCR link)
PCR Is a common lab technique used to make many copies of a DNA sample to work with or store in a laboratory. To learn more about it, check out these links!
https://www.youtube.com/watch?v=iQsu3Kz9NYo
https://media.hhmi.org/biointeractive/vlabs/bacterial_id/index.html (interactive PCR link)
Protein Purification
Protein purification is where you isolate protein from cellular matter. Protein purification is used because we need to isolate proteins in order to better understand the function and characterize a protein. There are many types of protein purification, affinity chromatography using Nickel being the most common. To learn more, check out these resources:
Protein purification is where you isolate protein from cellular matter. Protein purification is used because we need to isolate proteins in order to better understand the function and characterize a protein. There are many types of protein purification, affinity chromatography using Nickel being the most common. To learn more, check out these resources:
- Thermo Fisher Website on Protein Purification: https://www.thermofisher.com/us/en/home/references/protein-analysis- guide/affinity-chromatography/what-is-protein-purification.html
Very advanced NCBI link on protein purifcation: https://www.ncbi.nlm.nih.gov/books/NBK22410/
Gel Electrophoresis
Gel electrophoresis is a lab technique used to separate DNA, RNA and proteins over a charged gradient. The two types of gel electrophoresis that we use in the lab are agarose gel electrophoresis for DNA and polyacrylamide gel electrophoresis (SDS-PAGE) for proteins. Check out the links below!
Agarose Gel Electrophoresis: https://www.youtube.com/watch?v=saJIWFUGEbw Protein Gel Electrophoresis (SDS PAGE): https://www.youtube.com/watch?v=eaETFKXtNRA
Interactive: https://learn.genetics.utah.edu/content/labs/gel/
Gel electrophoresis is a lab technique used to separate DNA, RNA and proteins over a charged gradient. The two types of gel electrophoresis that we use in the lab are agarose gel electrophoresis for DNA and polyacrylamide gel electrophoresis (SDS-PAGE) for proteins. Check out the links below!
Agarose Gel Electrophoresis: https://www.youtube.com/watch?v=saJIWFUGEbw Protein Gel Electrophoresis (SDS PAGE): https://www.youtube.com/watch?v=eaETFKXtNRA
Interactive: https://learn.genetics.utah.edu/content/labs/gel/
Bacterial Transformation
Bacterial Transformation is a horizontal gene transfer where bacterial plasmids (usually E. coli) take up foreign genetic material from the environment (this "foreign genetic material" is our DNA of interest in the lab).
Khan Academy video overview: https://www.khanacademy.org/science/biology/biotech-dna-technology/dna-cloning-tutorial/a/overview-dna-cloning
Interactive Lab: http://labcenter.dnalc.org/labs/transformation/transformation_h.htm
Bacterial Transformation is a horizontal gene transfer where bacterial plasmids (usually E. coli) take up foreign genetic material from the environment (this "foreign genetic material" is our DNA of interest in the lab).
Khan Academy video overview: https://www.khanacademy.org/science/biology/biotech-dna-technology/dna-cloning-tutorial/a/overview-dna-cloning
Interactive Lab: http://labcenter.dnalc.org/labs/transformation/transformation_h.htm
Plasmid Miniprep
Plasmid Miniprep (plasmid purification) is used to purify and prepare your plasmid DNA for future experiments. In the lab, we follow the New England BioLabs protocol using a Monarch Plasmid Miniprep kit.
Miniprep use: https://www.youtube.com/watch?v=7Xgy5_i6iOc
NEB protocol: https://www.neb.com/products/nucleic-acid-purification/plasmid-purification/plasmid-purification
Plasmid Miniprep (plasmid purification) is used to purify and prepare your plasmid DNA for future experiments. In the lab, we follow the New England BioLabs protocol using a Monarch Plasmid Miniprep kit.
Miniprep use: https://www.youtube.com/watch?v=7Xgy5_i6iOc
NEB protocol: https://www.neb.com/products/nucleic-acid-purification/plasmid-purification/plasmid-purification
IPTG Induction
IPTG Induction of Cell Cultures is used to induce your recombinant DNA in order for your DNA to transcribe the target genes in your DNA. The mechanism of this is similar to the concept of the lac operon! For more information on this, check out:
Article: https://www.goldbio.com/articles/article/a-deep-dive-into-iptg-induction
Lac operon concept (will help to explain the induction of DNA expression): https://www.khanacademy.org/science/biology/gene-regulation/gene-regulation-in-bacteria/v/lac-operon
IPTG Induction of Cell Cultures is used to induce your recombinant DNA in order for your DNA to transcribe the target genes in your DNA. The mechanism of this is similar to the concept of the lac operon! For more information on this, check out:
Article: https://www.goldbio.com/articles/article/a-deep-dive-into-iptg-induction
Lac operon concept (will help to explain the induction of DNA expression): https://www.khanacademy.org/science/biology/gene-regulation/gene-regulation-in-bacteria/v/lac-operon
Electrophoretic Mobility Shift Assay (EMSA)--ADVANCED Technique
EMSA is a lab technique is a common electrophoresis technique used to study protein DNA or RNA interactions.
Information on the technique is below, though this is an advanced technique:
Video: https://www.youtube.com/watch?v=73fip8IC3Zo
Webpage: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2757439/
EMSA is a lab technique is a common electrophoresis technique used to study protein DNA or RNA interactions.
Information on the technique is below, though this is an advanced technique:
Video: https://www.youtube.com/watch?v=73fip8IC3Zo
Webpage: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2757439/
Northern, Southern and Western Blot--ADVANCED Technique
Northern, Southern and Western Blots are laboratory methods used to detect RNA, DNA and protein activity, respectively. You will likely not have to employ this technique in the lab, but in case you want to learn about it:
Northern Blot: https://www.youtube.com/watch?v=Zy7eKoJoFCs
Southern Blot: https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/southern-blot
Western Blot: https://www.youtube.com/watch?v=Ll_7z4YS2Ak
Northern, Southern and Western Blots are laboratory methods used to detect RNA, DNA and protein activity, respectively. You will likely not have to employ this technique in the lab, but in case you want to learn about it:
Northern Blot: https://www.youtube.com/watch?v=Zy7eKoJoFCs
Southern Blot: https://www.khanacademy.org/test-prep/mcat/biomolecules/dna-technology/v/southern-blot
Western Blot: https://www.youtube.com/watch?v=Ll_7z4YS2Ak