miniprep

목차

1. Title
2. Object
3. Introduction
4. Material & Methods
5. Result
6. Discussion
7. Reference

본문내용

1. Title : Miniprep

2. Object
Increase the amount of purified DNA using alkaline lysis method that seperates DNA according to its size

3. Introduction

[Alkaline lysis method]

Alkaline lysis is a method used in molecular biology to break cells open to isolate plasmid DNA or other cell components such as proteins. Bacteria containing the plasmid of interest is first grown, then lysed with a strong alkaline buffer consisting of a detergent sodium dodecyl sulfate (SDS) and a strong base sodium hydroxide. The detergent breaks the membrane`s phospholipid bilayer and the alkali denatures proteins involved in maintaining the structure of the cell membrane. Through a series of steps involving agitation, precipitation, centrifugation, and the removal of supernatant, cellular debris is removed and the plasmid is isolated and purified. The protocol may vary slightly from lab to lab.

<중 략>

From the result, we couldn`t get any DNA bands from the electrophoresis data. G4,G2 data show to be good as band is approximately 300, 600, 700, 1000, 2500 bp. Why G5 result couldn`t get any bands like G1? I searched the internet website to get reason for that. There is same questions and answers written by lab researchers. I attached solutions from well trained researchers.
First answer is checking the below things.

참고 자료

http://en.wikipedia.org/wiki/Alkaline_lysis
http://www.ehow.com/facts_7554689_lysis-protocol.html
http://public.wsu.edu/~kahn_sci/Flow/E2-QIAprep_Miniprep_Handbook.pdf
http://systemsbiology.yonsei.ac.kr/sub/catalog.php?CatNo=63&OutLine=menu1outline
http://www.scientistsolutions.com/t5252-miniprep+problems.html