Lab review

Lab review - Re-grade for Midterm 3:Students in Tuesday and...

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Unformatted text preview: Re-grade for Midterm 3:Students in Tuesday and Wednesday labs: submit re-grades to anyBISC 320 TA between 10-11 am on Friday, Nov 30th in Biospace (ZHS 360A)Students in Thursday and Friday labs: submit re-grades to any BISC 320 TA between 1-2 pm on Monday, Dec 3 in Biospace (ZHS 360A)Lab 1Lab 1Spectrophotometric Analysis of Nucleic Acids Topics:How to quantify nucleic acids concentration How to assess purity of nucleic acidsHow to calculate and perform dilutionsHow to operate spectrophotometerUltraviolet (UV, 200-400 nm) spectrophotometry can be used to - determine the quantity of nucleic acids (quantitative) - estimate nucleic acid purity (qualitative)- amount of energy absorbance is proportional to [substance]Absorption is primarily due to the aromatic rings. Purine Pyrimidine Spectrophotometric Analysis of Nucleic Acids Differences in Nucleotides of DNA and RNAIn DNA, the 2 hydroxylgroup of ribose is missing.Uracil is a Thymine without the methylgroup Temperature (C)Absorbance at 260 nmds DNAss DNA (weaker interaction between base pairs higher abs)DNA Melting CurveAromatic bases are stacked in the hydrophobic coreA260/ A280ratio indication of protein contamination(1.8~1.9 for DNA and 1.9~2.0 for RNA)A230 Phenol or Urea contaminationA325 Dirty cuvetesA 260/A 230 ratio:a secondary measure of nucleic acid purity. The 260/230 values for pure nucleic acid are often higher than the respective 260/280 values. Expected 260/230 values are commonly in the range of 2.0-2.2. Qualitative Measurement of Nucleic Acid 1 :5 0 1 :1 0 0 1 :2 0 0 1 :8 0 1 :2 5 0 1 :4 0 0 S a m p le 1 (l) S a m p le 2 (l) H2O (l) H2O (l) F in a l Vo lu m e (m l) 1 .0 1 .0 1 .0 F in a l Vo lu m e (m l) 1 .0 1 .0 1 .0 1 :5 0 1 :1 0 0 1 :2 0 0 1 :8 0 1 :2 5 0 1 :4 0 0...
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This note was uploaded on 09/17/2009 for the course BISC 320L taught by Professor Baker,aparicio during the Fall '07 term at USC.

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Lab review - Re-grade for Midterm 3:Students in Tuesday and...

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