Making a Recombinant DNA, Step by Step

Name: Making a Recombinant DNA, Step by Step
Author: StemPy Lab

A gene is a few thousand letters inside a genome of millions. How do you find it, copy it and put it into a cell? You follow the steps in order. You isolate the DNA, cut it and read the pattern on a gel. You copy one gene a billion times by PCR. Then you join the gene to a vector, put it into a host and pick out the cells that took it. It serves the Indian syllabus (NCERT, NEET) and the matching topic of A-Level, IB and AP.

3 lessons, about 1 h 30 min3 simulations11 problems3 practice questionsPython in every lesson

Created by StemPy Lab

Course outline (3 lessons)

Lesson 1about 30 min
Isolating the DNA, cutting it and reading the gel
To clone a gene you must first get the DNA out of a cell. Enzymes break the wall, and other enzymes remove the RNA and the proteins. Chilled ethanol makes the DNA appear. Restriction enzymes then cut it, and gel electrophoresis separates the fragments by size, so that the right piece can be cut out of the gel.
1 simulation1 practice question1 Python program4 problems1 Python lab
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Lesson 2about 30 min
Amplifying a gene by PCR
The polymerase chain reaction copies a chosen piece of DNA again and again. It needs two primers, the four nucleotides and a polymerase that survives heat. Each cycle has three steps: denaturation, annealing and extension. The number of molecules doubles in each ideal cycle, so 30 cycles give about a billion copies.
1 simulation1 practice question1 Python program4 problems1 Python lab
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Lesson 3about 30 min
Joining, transferring and selecting the recombinants
The cut vector and the gene are joined by ligase, and the recombinant DNA is put into a host. A bacterium must be made competent, by calcium and a heat shock. Animal and plant cells take DNA by microinjection, a gene gun or a disarmed vector. Cells that took the DNA are found by a selectable marker. Recombinants are found by insertional inactivation.
1 simulation1 practice question1 Python program3 problems1 question set1 note
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Making a Recombinant DNA: Practice Paper

20 questions · 15 min

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Making a Recombinant DNA, Step by Step

Course outline (3 lessons)

Isolating the DNA, cutting it and reading the gel

Amplifying a gene by PCR

Joining, transferring and selecting the recombinants

Test Your Learning

Your teacher

Discussion (0)