Week_5_handout

Week_5_handout - Bi 1x Spring 2010 Week 5 o Agarose gel...

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Page 1 of 2 Bi 1x, Spring 2010 Week 5 o Agarose gel electrophoresis Materials Your digestion reactions from last week 6X DNA loading dye 1 kb and 100 bp ladders (pre-stained) HindIII digested lambda DNA (pre-stained) pZE21-lacZ DNA (pre-stained) Your PCR reactions Background Agarose gel electrophoresis is a powerful separation method frequently used to analyze DNA fragments generated by restriction enzymes. The agarose gel consists of microscopic pores that act as a molecular sieve. Samples of DNA are loaded into wells made in the gel during casting, as shown in the figure below. http://ocw.mit.edu/OcwWeb/Biological-Engineering/20-109Fall-2007/Labs/detail/mod1_2.htm Since DNA has a strong negative charge at neutral pH, it migrates through the gel towards the positive electrode during electrophoresis. The DNA molecules are separated in the gel according to their size and shape. Linear DNA molecules are separated according to their size. The smaller the linear fragment, the faster it migrates. If the size of two fragments are similar or identical, they will
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