Synthesis of low molecular weight DNA ladder / Nurul Farhana Kamarulzaman

DNA ladder is an important tool in Gel Electrophoresis to determine the size of DNA fragment of (Polymerase Chain Reaction) PCR products. It has two types which is low molecular weight and high molecular weight DNA ladder. The objective of this research is to synthesize low molecular weight DNA ladd...

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Bibliographic Details
Main Author: Kamarulzaman, Nurul Farhana
Format: Thesis
Language:English
Published: 2009
Subjects:
Online Access:https://ir.uitm.edu.my/id/eprint/105840/1/105840.PDF
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Summary:DNA ladder is an important tool in Gel Electrophoresis to determine the size of DNA fragment of (Polymerase Chain Reaction) PCR products. It has two types which is low molecular weight and high molecular weight DNA ladder. The objective of this research is to synthesize low molecular weight DNA ladder. It is also known as 100bp DNA ladder. There are several methods to synthesize low molecular weight DNA ladder. In this research, the method used is by PCR amplification of lambda phage DNA. It is done by designing PCR primer for each length of I OObp ladder. Then,PCR process is done and continued by running gel electrophoresis. After that, the gel is screened through UV transilluminator. As a result, there are several successful PCR product which are 200bp, 300bp, 400bp,600bp, 700bp, 800bp and 1000bp. The remaining shows failure of PCR process although the procedures are repeated for several times. The reason of failure is maybe due to contamination of reagents used. Although there are failure for some of the PCR amplification, but the method to synthesize of low molecular weight DNA ladder using PCR amplification is really useful to reduce laboratory cost compared buying it from the commercial DNA ladder.