Constructing a DNA ladder Range for Lambda Phage by multiplex PCR
Abstract
Background and Objectives: DNA ladder contains DNA fragments of different length but with known size, used to determine the size of unknown DNA molecules. Different DNA ladders are available for expected DNA length. Conserved sequences were selected for design of primers to generate DNA fragments of known specific size.
Materials and Methods: In this study, we describe a method by which DNA ladder was prepared based on multiplex PCR technique. Different lengths of DNA fragments were amplified using the primers designed according to the 1216-2136 sequence extent of lambda phage DNA. Target DNA fragments were amplified using multiplex PCR and extracted.
Results: The results showed an amplified lambda phage DNA at particular target sites by using 1 forward and 6 different reverse primers (for 100, 200, 400, 600, 800, 1000bp) for the successful amplification.
Conclusion: This method would be more cost effective than commercial DNA molecular weight markers.
Van der Vliet GM, Hermans CJ, Klatser PR. Simple colorimetric microtiter plate hybridization assay for detection of amplified Mycobacterium leprae DNA. J Clin Microbio 1993; 31: 665-670.
Benson SA, Taylor RT. A rapid small-scale procedure for isolation of phage lambda DNA. Biotechniques 1984; 2: 126-127.
Sambrook J (2001). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New Yark.
Hartley J. L (2006). Nucleic acid marker ladder for esti- mating mass. US Patent no. 7, 132, 520.
Kornreich R, Desnick RJ. Fabry diseases: A gene by multiplex PCR amplification. Hum Mutat 1993; 2: 108-111.
| Files | ||
| Issue | Vol 2 No 4 (2010) | |
| Section | Articles | |
| Keywords | ||
| Lambda phage DNA DNA ladder Multiplex PCR | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
