MSC IV SEMESTER_DNA Profiling - CE.pdf

DNA Profiling - CE

1. DNA Profiling

8. https://strbase-archive.nist.gov/index.htm

24. Glass capillaries with viscous polymer

30. ELECTROKINETIC
INJECTION

32. Sample Preparation
PCR products dilution
1 in 10 with deionized formamide
Reduce salt conc, denaturation of dsDNA, aid in
electrokinetic injection process
“snap-cooling” : 95 degree C for several minutes
followed by rapid cooling to around 4 degree Celsius
Hi-Di formamide
(Applied Biosystems,
Foster City, CA)

33. DNA SEPARATION
Polymer Buffer
Capillary tube
Electric field /
Voltage 15,000
volts
Ph 5-8

34. POLYMER BUFFER
POP-4 and POP-6, POP-7where “POP”
stands for Performance Optimized
Polymer.
100mmol/L TAPS and 1mmol/L EDTA,
adjusted to pH 8.0 with NaOH
TAPS is short for N-tris-
(hydroxymethyl)-methyl-3-
aminopropane-sulfonic acid.
These polymer solutions are 4% and
6% concentrations of linear, uncross-
linked dimethyl polyacrylamide,
respectively
TAPS is used instead of Trisborate-
EDTA (TBE) since TBE is temperature
and pH-sensitive.
A high concentration of urea buffer additives such as formamide,
urea, and 2-pyrrolidinone
POP-4 consists of 4% linear
dimethylacrylamide, 8mol/L urea,
5% 2-pyrrolidinone
capillary run temperatures of 60oC

35. A number of factors
influence resolution in a
CE system
capillary length
separation voltage
polymer concentratioN

52. Comments
on Sample
Preparation

53. This Photoby Unknown Author is licensed under CC BY-SA