Molecular Biotechnology Lab Report

INTRODUCTION –

Rapid advancements in the field of Biotechnology require understanding of underlying principles of how the basic techniques are involved in Molecular Biology. Expertise on basic Molecular Biological techniques is considered essential for pursuing advanced level research in this exciting field. In order to keep high academic standard, it is mandatory to know tools and techniques used for exploration of living system. The major objective during the summer internship was to deal with the molecular biology techniques. Molecular biology is the study of science that deals with the molecular basis of biological activity. Many advances

DNA extraction of Litsea glutinosa,

2. Purification and quantitation,

3. Agarose gel electrophoresis, &

4. Polymerase chain reaction. For genomic DNA extraction protocol of the Deshmukh et al. (2007) will be used. This protocol is based on the SDS and suitable for species having higher amounts of secondary metabolites, gums and resins. Genomic DNA of Litsea glutinosa will be extracted. The DNA concentration plays an very important role in many applications in molecular biology like complete digestion of DNA by restriction enzymes and amplification of target DNA by polymerase chain reaction. To measure the quantity of DNA, It is generally carried out mainly by 2 methods- 1. Spectrophotometric and

2. Ethidium bromide fluorescence. In Spectrophotometric method, an absorbance ratio of 260 nm and280 nm gives an estimate of the purity of the solution.Pure DNA and RNA solutions have optical density 260 & 280 nm which have values of 1.8 and 2.0, respectively. Therefore a ratio higher than 1.8 indicates RNA contamination and a ratio lower than 1.8 indicates protein

The species from different locations was taken to examine each and every aspect related to the plants and its functional properties

1. DNA EXTRACTION -

The protocol of Deshmukh et al. (2007) was followed.This protocol is based on SDS and suitable for this plant.The leaves of Litsea glutinosa are crushed with the help of liquid nitrogen for the better results. This plant contains secondary metabolites and high amount of gums and resins.The genomic DNA isolation needs to separate total DNA and RNA, protein, lipid etc..Initially the cell membrane must be disrupted for successful release of DNA in extraction buffer.SDS disrupt the cell membrane. 2. PURIFICATION AND QUANTITATION OF EXTRACTED DNA-

In the Spectrophotometric method, an absorbance ratio of 260 and 280 nm estimate the purity of DNA. The A260/A280 RATIO around 1.8 (1.75-1.85) indicates best quality DNA.As per expectation ,after RNAs treatment the yield of DNA was slightly declined in methods other than Deshmukh et al. (2007) as in this method yield is