The % of Transformation Efficiency = Practical yield / Theorical yield * 100.
The practical yield = 10^6/ ug and
the theoritical yield = 10^8/ ug.
So, The TE = 10^6/10^8 * 100 = 1.
so, the answer is 1.
4. You have prepared a batch of competent E coli cells and found its transformation efficiency...
You add (1.04x10^0) ng of plasmid DNA in 50 µL to 100 µL of competent E. coli cells and incubate the mixture on ice for 20 min. Following a heat-shock treatment, the cells are incubated at 37 C for 30 min. At the end of the incubation you spread (1.2000x10^2) µL of the transformation mix on an agar plate and incubate the plate overnight at 37 C. The next morning you observe (6.8400x10^2) colonies on the plate, each colony having...
Bacterial cells, such as E. coli, are surrounded by a fluid plasma membrane and enclosed by a rigid cell wall that protects the cell from damage. You will recall that there is no nuclear envelope in prokaryotic cells, but rather a “nucleoid region” with a single chromosome that is continuous (circular), not linear, as in eukaryotes. All of the genes required for basic survival and reproduction are found in the main chromosome. In addition to the main chromosomes, some DNA...
1. Fill in the table above with what you observe on your plates. 2. Bacterial transformation occurred on which agar plate (s)? What evidence do you have that the bacteria were transformed here? 3. Which plates have glowing growth? Explain what causes bacteria to glow. II. Transformation of E. coli with Plasmid DNA (PGLO) 1. Three LB (Luria Broth) agar plates are obtained. The plates contain: • Plate A: LB-Agar/Ampicillin/Arabinose • Plate B: LB-Agar/Ampicillin • Plate C: LB-Agar 2. Three...
The starting concentration of the plasmid DNA is 100 ngul and you added 2μ1 to your cells. The cells were diluted to one ml prior to plating. First, determine how many transformants you would have if you plated the entire 1 ml of cell:s. Remember, you only plated half or your transformation reaction on plates 1 and 2. Add the number of transformants on plates 1 and 2, and then multiply by 2 for the total number transformants. Record that...
Suppose that you carried out a Bacterial transformation of E. coli HA101 with pGLO plasmid experiment in the lab. During the experiment plates with bacteria were inoculated from +GLO and -GLO microfuge tubes (LB (-) plate, LB/amp (-) plate, LB/amp (+) plate, and LB/amp/ara (+) plate). 14) Explain what kinds of bacterial growth you will find on each of 4 plates after incubation ((LB (-) plate, LB/amp (-) plate, LB/amp (+) plate, and LB/amp/ara (+) plate): under normal light and...
PLEASE SHOW ALL WORK IF POSSIBLE! THANK YOU IN ADVANCE. 1) SOC media is 2% tryptone, 0.5% yeast extract, 10 mM NaCL (F.W. 58), 2.5 mM KCl (F.W. 74.5), 10 mM MgSO4 (F.W. 246.5), 10 mM MgCl2 (F.W. 203.3), 20 mM glucose (F.W. 180.16). Determine the amounts of each to make 500 mL of SOC. 2) Determine the number of moles in 0.05 μg of 300 bp insert DNA. There are 660 g/mol-bp. What is the molar ration of insert...
You have a strain of E. coli which is ade- (adenine minus, meaning unable to make adenine on its own). In the biosynthesis of adenine, there are five steps as follows: In the biosynthetic pathway shown above, the ‘precursor’ is the starting material. Enzyme 1 converts this precursor to compound 1 (intermediate 1), enzyme 2 then acts on compound 1 to give rise to compound 2 (intermediate 2), so on and so forth, until enzyme 5 gives rise to the...
LAB Genetic Engineering of Bacteria Problem Is it possible to transfer the allele for resistance to the antibiotic ampicillin into a bacterial cell? Objectives After completing this lab, the student will be able to: 1. Demonstrate micropipetting and sterile pipetting techniques for handling and transferring bacteria and plasmid DNA. 2. Maintain sterile conditions for culturing bacterial cells. 3. Inoculate bacteria into flasks, culture tubes, or agar plates. 4. Culture isolated individual colonies from an agar plate to form genetically identical...
As a student project, you have isolated six new mutant strains of E. coli with altered behavior of the lactose operon. The strains are listed in the table below, together with their phenotypes (with regard to significant ?-galactosidase synthesis) in three specific situations. Columns 1 and 2 present the phenotypes of each mutant haploid strain. In column 1, the mutant is in an otherwise wild-type genome. In column 2, the genome also carries a nonsense-suppressor mutation (that is not present...
LAB17 Genetic Engineering of Bacteria Problem Is it possible to transfer the allele for resistance to the antibiotic ampicillin into a bacterial cell? Objectives After completing this lab, the student will be able to: 1. Demonstrate micropipetting and sterile pipetting techniques for handling and transferring bacteria and plasmid DNA. 2. Maintain sterile conditions for culturing bacterial cells. 3. Inoculate bacteria into flasks, culture tubes, or agar plates. 4. Culture isolated individual colonies from an agar plate to form genetically identical...