Biochemistry 2280A Lecture Notes - Palindromic Sequence, Autoradiograph, Enzyme Kinetics

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It is important to recognize that most applications of molecular biology use chemical quantities of the dna. In a dna sequencing reaction you generally add ~100 ng of dna. Dna is 3000bp, approximately how many moles of template are added: 5x10-14 moles, 1 mole of a dna of 3000bp has a mass of: 3000 x 660g/bp ~2 x 106 grams in one mole. 100 ng has 5 x 10-14 moles. That is there are a lot of molecules of template dna in a sequencing reaction. Restriction enzymes are usually type ii enzymes, which show specific recognition and specific cleavage of dna. There are about 300 type ii restriction enzymes now known. They are site-specific dna binding proteins, which in general recognize and cut palindromic sequences most commonly of 4, 6 and 8 bases. It is symmetric on both strands (leading and lagging) After cleaving you get 5" overhangs on either end (single stranded) The 5" end is what is overhanging.

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