AdministratorHow big is the pGLO plasmid?
5371 bp
Size: the pGLO plasmid is 5371 bp. The GFP coding region is about 714 bp, and the GFP protein is about 26 kD, with 238 amino acids.
What is the size of pGLO how many base pairs is it?
5371 base pairs
This allows for more precise determination of the size of the DNA fragments. In this lab experiment, you will start with two different plasmids that are close to the same size (pGLO is 5371 base pairs long, and pARO180 is 5460 bp).
What is +pGLO LB amp?
+pglo LB AMP ARA: GLOWing bacteria that are transformed , large in size because of the arabinose added. SATELLITE colonies. These are colonies that don’t glow but grow. -pglo LB/ AMP: no growth, this is b/c no bla gene is present.
How do you measure the size of a plasmid?
You can get an approximate size by cutting it with a restriction enzyme and seeing what size fragments are produced. It might take testing with a few different enzymes to find one that cuts into only a few fragments, so you can add the fragments sizes up to get the total.
How do you determine the size of a fragment from a plasmid?
To calculate the fragment size we simply need to subtract the bp difference between the two REs: 5198 bp – 5070 bp = 128 bp. DNA fragment 2 – a large fragment (and our desired DNA fragment as it contains all the features of the plasmid.
How many band’s should be expected if the uncut pGLO plasmid is loaded as a sample on the gel?
3 bands
When uncut plasmid DNA is isolated and run on an agarose gel, you are likely to see 3 bands.
Is pGLO a plasmid?
Bio-Rad’s pGLO plasmid contains DNA sequences that enable its replication and expression of the fluorescent trait (phenotype) in bacteria following transformation.
What is the purpose of the LB amp ARA on an agar plate?
What is the purpose of the LB/amp/ara agar plate? It is a selective medium with an antibiotic which prevents growth of non-resistant cells and a sugar that allows for the expression of a plasmid gene.
What is LB amp plate?
Application. LB agar plate is used to culture bacteria in microbiology research. These plates are commonly used during cloning experiments to amplify antibiotic-resistant, competent bacteria. The addition of agar to LB medium helps form a gel that allows bacteria to grow.
What does LB mean in pGLO lab?
LB: luria broth-nutrients. Amp: the antibiotic ampicillin. Ara: the sugar arabinose. +pGLO: contains transformed bacteria. -pGLO: contains non-transformed bacteria.
What is pGLO plasmid made of?
The pGLO plasmid contains both the promoter (pBAD) and araC gene, but araB, araA, and araD have been replaced by the single gene that codes for GFP, which serves as a reporter gene.
What is in Biobio-rad’s pGLO plasmid?
Bio-Rad’s pGLO plasmid contains DNA sequences that enable its replication and expression of the fluorescent trait (phenotype) in bacteria following transformation. The essential sequences include the following:
What is a pGLO plasmid and GFP kit?
Bio-Rad Explorer pGLO Plasmid and GFP Kits use the pGLO plasmid, which contains the GFP gene, to enable hands-on learning about the central dogma, gene expression and regulation, bacterial transformation, protein separation, and the biomanufacturing process.
What is a pGLO lab kit?
pGLO Lab Kits utilize Bio-Rad’s pGLO plasmid, which encodes a green fluorescent protein (GFP), to enable instructors to give students a hands-on introduction to transformation, cloning, protein chromatography, and electrophoresis techniques. Apply a three-dimensional approach to the classic pGLO bacterial transformation activity.
Why do bacteria transformed with pGLO show green fluorescence?
Gene expression and gene regulation — bacteria transformed with pGLO show green fluorescence only when they are grown in the presence of arabinose. Use this experiment to illustrate that not all genes are expressed at the same time and that environmental factors control when gene expression takes place