review mehmet - SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC...

Info icon This preview shows pages 1–10. Sign up to view the full content.

View Full Document Right Arrow Icon
SPECTROPHOTOMETRIC ANALYSIS OF NUCLEIC ACIDS
Image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Objectives Learn how to do dilutions Use a spectrophotometer to calculate concentrations of nucleic acids Use absorbance data to figure out the purity of nucleic acids
Image of page 2
Spectroscopy Nucleic acids absorb light Best absorbance at 260nm
Image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Absorbance and Concentration Different molecules have different absorbance properties Absorbance reading of 1.0: dsDNA: 50 ug/ml ssDNA or RNA: 40 ug/ml Single stranded oligonucleotides: 33 ug/ml
Image of page 4
Absorbance and purity 280nm: absorbance peak for proteins A260 / A280 ratio for purity 1.8 to 1.9 indicate high quality DNA. Low ratios indicate proteins. 1.9 to 2.0 for RNA 230nm: absorbance peak for phenol A260 / A230 ratio for purity. The lower the less pure. A325: contamination from cuvette. Should be 0
Image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Dilutions Example: 1:20 dilution Total volume is 20 parts. 1 part of it is the sample (stock). The remaining 19 parts is water. Say you want the total volume to be 1ml (1000ul) Volume of sample = 1000 / number of parts. 1000 / 20 = 50 ul How much water? 1000 – 50 = 950 ul
Image of page 6
DNA Electrophoresis Objective: to run electrophoresis of PCR amplified Alu insert to determine genotype for Alu insertion to perform Hardy-Weinberg analysis of class result
Image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Principle of Gel Electrophoresis
Image of page 8
Expected Results 941bp 641bp Alu -300bp intron intron Primer 1 Primer 2 +/+ : 941bp -/- : 641bp ( )
Image of page 9

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Image of page 10
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern