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Understanding Gel Electrophoresis: The DNA Amplification and Separation Process

In this educational resource, students will learn the process of gel electrophoresis through a series of informative videos. The process starts with obtaining a DNA sample, followed by amplification using Polymerase Chain Reaction (PCR). Students will see how restriction enzymes cut the DNA at specific sequences, creating fragments suitable for electrophoresis. The subsequent steps of preparing the gel, loading DNA samples, and staining the gel to visualize the results will be covered in detail.

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Understanding Gel Electrophoresis: The DNA Amplification and Separation Process

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Presentation Transcript

  1. Objectives: SWBAT outline the process of gel electrophoresis by watching several short videos of the process. Obtain a sample of DNA. PCR Polymerase Chain Reaction is used to amplify the amount of DNA. Use restriction enzymes to cut the DNA in pieces. • http://www.youtube.com/watch?v=RVhitK_b4gg • pcr polymerase chain reaction to amplify dna

  2. Restriction enzymes recognize a specific sequence of nucleotides and produce a double-stranded cut in the DNA. The recognition sequences usually vary between 4 and 8 nucleotides, and many of them are palindromic, meaning the base sequence reads the same backwards and forwards. Themirror-like palindrome is similar to those found in ordinary text, in which a sequence reads the same forward and backwards on a single strand of DNA strand, as in GTAATG. The inverted repeat palindrome is also a sequence that reads the same forward and backwards, but the forward and backward sequences are found in complementary DNA strands. EcoRI digestion produces "sticky" ends; usually a four base overlap that can be used to attach the sequences together with ligase enzyme.

  3. 4. The gel is prepared with wells (depressions that will hold the DNA). 5. A dye is added to the DNA to track the movement during the electrophoresis process. 6 . DNA is loaded into the wells using a micropipette. 7. The machine is started & DNA is drawn towards the positive end with smaller fragments travelling a greater distance. 8. The gel is stained to reveal the band patterns. http://www.youtube.com/watch?v=tTj8p05jAFM 2. loading the dna http://www.youtube.com/watch?v=8Afh_0IAfrQ 3. using a micropipette

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