BIO 615
Name: NUR LISMA RUHILA BT ALIAS
Group: AS201 5A
Experiment: GEL ELECTROPHORESIS OF EXTRACTED DNA 0.5% AGAROSE GEL
Group partners: 1) HALIMATUN SAADIAH BT MOHD BUSTAMAM 2) NUR FARHANA BT AHMAD SOPIAN 3) FATIN NUR ASYIQIN BT ABD TALIB 4) UMMU AFIQAH BT HASSAN 5) NABIHAH BT MD NAWAWI
Date of experiment: 8th October 2012
Date of submission: 15th October 2012
TITLE: GEL ELECTROPHORESIS OF EXTRACTED DNA 0.5% AGAROSE GEL
DATE: 8th OCTOBER 2012
OBJECTIVE
* To study measure the size of base pair of DNA
RESULT
Lane from extremely left:
Lane 1: Empty
Lane 2: Elute 2
Lane 3: Empty
Lane 4: Empty
Lane 5: Elute 1
Lane 6: Empty
Lane 7: Marker
Lane 8: Empty
1 2 3 4 5 6 7 8
The film paper showed the DNA band (Elute 1 and Elute 2) and marker gene (Hind III) which moves along the way from the negative pole to the positive pole of the electric field in agar medium. Each of the band mark by marker gene has its own base pair. It can be said that the DNA band for Elute 1and Elute 2 has the same base pair which are 23,130 bp with mass 477 μg/1μg.
DISCUSSION
Gel electrophoresis is a method used to separate DNA fragments according to its length using electric field and agar medium. Each band on the gel diagram represents base pairs of DNA. DNA band moves from the negative pole to the positive pole of the electric field as it is negatively charged due to the presence of phosphate group. The agarose gel acts as a matrix of tiny pores that allow small particles to move through it relatively quickly. As the fragments moves through the pores of agar medium, the shorter fragments move faster than the longer one. As a result, this method separates the DNA fragments according to their sizes. The nearest to the negative pole is the shortest fragment. In this experiment, HindIII digest of lambda DNA,
References: 1. Strege et al, (1991), Anion-exchange chromatography of DNA restriction fragments , Journal of Chromatography A, Retrieved October-9-2012 2. N.P Singh, R.E Stephens, (1991), Microgel electrophoresis: Sensitivity, mechanisms, and DNA electrostretching , Mutation Research/DNA Repair, Pages 167-175, Retrieved October-9-2012 3. Siguang Jiang, Xiaodong Chen, Minghua Liu, (2004), The pH stimulated reversible loading and release of a cationic dye in a layer-by-layer assembled DNA/PAH film, Journal of Colloid and Interface Science, Pages 396-403, Retrieved October-9-2012 4. Robert F.Weaver. Molecular Biology. (5th ed.). McGraw Hill International Edition, United States. 76-79pp 5. Keren DF, Gulbranson R, Ebrom SJ, (2004), False-negative urine protein electrophoresis by semiautomated gel electrophoresis, Clin Chem, Pages 396-403, Retrieved October -12-2012 6. Ganguly A, Rock MJ, Prockop DJ, (1993), Conformation-sensitive gel electrophoresis for rapid detection of single-base differences in double-stranded PCR products and DNA fragments: evidence for solvent-induced bends in DNA heteroduplexes. Proc Natl Acad Sci U S A, Pages 10325-10329, Retrieved October-12-2012 7. Finnila S, Hassinen IE, Ala-Kokko L, Majamaa K, (2000), Phylogenetic network of the mtDNA haplogroup U in Northern Finland based on sequence analysis of the complete coding region by conformation-sensitive gel electrophoresis. Am J Hum Genet, Pages 1017-1026, Retrieved October-12-2012 8. Mussap M, Ponchia S, Zaninotto M, et al, (2006), Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein. Clin Biochem, pages 152-159, Retrieved October-12-2012 9. Ehrich JHH, Wurster U, (1991), Differentiation of proteinurias with electrophoresis. Pediatr Nephrol, Pages 376-378, Retrieved October-12-2012 10. Le Bricon T, erlich D, Bengoufa D, et al, (1998), Sodium dodecyl sulfate-agarose gel electrophoresis of urinary proteins: application to multiple myeloma. Clin Chem, Pages 1191-1197, Retrieved October-12-2012