Lecture3mcb160Lf11

Lecture3mcb160Lf11 - EcoRIHindIII doubledigest...

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Note that EtBr intensity is propor0onal to mass of fragments….in the ladder of bands each fragment is present in equimolar ra0o but larger fragments fluoresce more brightly. Also note that bands that are very similar in size may not be resolved and may run as a “doublet” …the unusually bright intensity rela0ve to size indicates the presence of twice the expected mass (two fragments of the same size) EcoRI – HindIII double digest HindIII digest Lambda DNA restric0on digests
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1 2 3 4 5 6 7 8 9 10 Measure from bottom of well to bottom of band 1,6 supercoil p160L 2,7 EcoRI cut p160L 3,8 Lambda, Hind III 4,9 Lambda, EcoRI-HindIII 5, 10 Nicked circle p160L Lane Sample Sample gel Lab 3
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Distance unknown band migrated Determine size of unknown fragment
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Par$al Digests
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Star Ac$vity Star Activity can arise from a number of causes: 1. Too much glycerol: Final glycerol conc. should never exceed 5% in a digest. Remember that the enyzme stock is in 50% glycerol.
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This note was uploaded on 01/25/2012 for the course MCB 160L160L taught by Professor Venkatesansundaresan during the Fall '11 term at UC Davis.

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Lecture3mcb160Lf11 - EcoRIHindIII doubledigest...

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