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Chapter 20 Blog: Genetic Technology (Kimberley)

Page history last edited by KimberleyHausheer 12 years, 11 months ago

A. Chapter 20 Summary
1. Materials are obtained. You need to isolate a vector and a gene of interestt.  The gene of interest is the particulat gene that is going to be cloned.  A vector is a  small amount of DNA that  acts as the carrier for the gene of choice. Vectors can either be small commercially engineered plasmids or viral .  Viral vectors are when you insert the gne of choice into the genetic material of the virus.
2. Create a Recombinant Vector. A recombinant vector is when a vector s  successfully takes up a piece of DNA.  First, resriction enzymes cut  the chromosomal DNA into many small fragments.  They cut at places where they find palindrominc sequewnceces.  Restriction enzymes also cut the vector DNA.  Then the fragments of DNA join into the open vector and form hydrogen bonds. DNA ligase permanently seals the fragment iinto the vector therefore creating a recombinant vector.
3. Replicating Recombinant Vectors.  Bacteria cells are treated so that they take up DNA molecules. If they do take up a vector than it will automaticlalyrepliate whenever the cell divides.

Gel electrophoresis is a technique that is used in  labs to separate macromolecules, such as DNA and proteins on a gel according to size , charge, or mass, using an electrical current. Samples are loaded into wells on the top of the gel. Then an electrical current is applied causing the molecules to pull towards the bottom of the gel. Because the gel is porous smaller molecules will be able to get through it easier and will be closer to the bottom than larger molecules. The fragments are than stained and examined.

At the end of the cloning experiment all of the bacterial cells are plated. TAll of the cells on the plate are considered the DNA library for that experiment. It consists of all of the cells that took up different recominant vectors that each have different fragments of DNA  Two types of DNA libraries are commonly mde. Genomic libraries are made of vectors containing fragments of chromosomal DNA. cDNA libraries are made of vectors containing complementary DNA fragments which are fragments of mRNA. cDNA libraries are preferred becaue mRNA does not have any introns.

B. Useful Materials

Comments (1)

Derek Weber said

at 1:55 am on Apr 2, 2011

No materials.

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