SBI4U: Biotechnology Tools and Techniques What is biotechnology? Applied biology genetics; molecular biology; microbiology; biochemistry Uses living organisms and their components to create “______________________” industry, agriculture, medicine Involves _____________________________ Manipulating DNA Recombinant DNA: A Fragment of DNA composed of ___________________________________ Biotechnology Tools and Techniques 1. Restriction Endonucleases 2. Methylases 3. Ligase 4. Gel Electrophoresis To join 2 DNA Sequences you need scissors and glue, but where can we find these items in a cell??? Restriction Endonucleases aka ________________________________ “molecular scissors” What do they do? recognize ______________________________________, and then ________ the double-stranded _____________________________ Function: Crude immune system in _______________ Cleaves _____ DNA into fragment Host DNA is methylated: RE knows not to cleave it Recognition site: the sequence ____________________________________ Characteristics:
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SBI4U: Biotechnology Tools and Techniques
What is biotechnology?Applied biology
genetics; molecular biology; microbiology; biochemistryUses living organisms and their components to create “______________________”
Can be used to join fragments that have __________________________________
_________________________ bond Most frequently used: ____________________
Gel Electrophoresis Method of separating DNA fragments Used in genetic engineering to isolate
____________________________ RE may cut at several sites. Want to make sure the ________________
__________________________________
Useful properties of DNA1. DNA is __________________________
(phosphate groups)2. Charge-to-mass ratio of all nucleotides is
__________________________
Principles of Gel Electorphoresis Separates DNA fragments based __________________ Involves forcing DNA fragments ____________________________________ Matrix acts like a sieve – has pores through which ________________________ Fragments will migrate through the gel at a rate that is ______________________________
to logarithm of their size Smaller fragments will ________________________ Larger fragments will _________________________
Producing Recombinant DNA
Procedure1. DNA is cleaved into ________________________. Depending on the cut sites, the fragments
will be different sizes.2. The sample of DNA is loaded into _____________________ within the gel matrix.3. A charge is applied across the gel: Negative at the sample end; positive at the opposite.4. DNA fragments will ___________________________________________
Depending on fragment size, migration rates will vary
Sizing the Bands A “____________________” of fragments of known sizes is run alongside samples Compare samples to ___________________________________
Visualizing the DNA Stain with ___________________________ Ethidium bromide inserts itself into the DNA backbone Fluoresces under ______________________
Obtaining the desired fragment Literally __________________________ Purify to obtain the fragment