Pair SW on RECOMBINANT DNA TECHNOLOGY – an online copy with clickable links is available at https://bayo2pisay.wordpress.com . Answer on intermediate pad paper.
Define the following terms using any reputable website:
1. recombinant DNA
2. cloned DNA
4. genetic engineering
Basic DNA technology techniques – Access the given sites and answer the questions that follow.
1. Restriction enzymes – http://highered.mcgraw-hill.com/olc/dl/120078/bio37.swf.
a) What are restriction enzymes/endonucleases?
b) Give an example and the sequence it identifies.
c) Give an application of these tools in genetic engineering.
2. DNA recombination and gene cloning http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html.
a) What are plasmids?
b) Draw a flowchart describing how foreign DNA is inserted into bacterial plasmids and how a whole colony of identical daughter cells can result. Be sure to include the terms vector, restriction enzyme, foreign DNA, ligase, sticky ends, daughter cell, colony, and clone.
c) In the example given, how do researchers ensure that only the host cell with the DNA sequence of interest will be cloned?
a) Why can’t eukaryotic DNA be cloned directy into prokaryotic cells to make useful proteins?
b) How does reverse transcriptase and the creation of cDNA solve problem 3a?
a) What is PCR?
b) Draw a flowchart describing how a cycle of PCR works. Be sure to include the following terms: target DNA, primers, nucleotides, Taq polymerase, denature, anneal, polymerize/extend.
c) After 25 cycles (which takes around 2 hours), how many copies of the target DNA are made?
5. Gel electrophoresis – http://www.dnalc.org/resources/animations/gelelectrophoresis.html
a) What is gel electrophoresis used for?
b) How is DNA separated in gel electrophoresis? Include the charge of DNA and the structure of the gel and the size of the fragments in your answer.
a) What method is of DNA sequencing is shown in this animation?
b) Draw a flowchart describing how a fragment of DNA is sequenced using this method.