Molecular Biology Lab Report Example | Topics and Well Written Essays - 1750 words

Molecular biological science - Lab Report ExampleDpn I and Fse I together fragments of 0.5 kb, 1.1 kb, 1.6 kb and 2.3 kbDpn I, Eag I and Fse I together fragments of 0.3 kb, 0.5 kb, 0.6 kb, 1.0 kb, 1.1 kb and 2.0kba) How many restriction sites be on that point for severally enzyme What, if any, are the strange restriction sites on this plasmidAns. Dpn I = 3, Eag I = 2, Fse I = No RS. There are remarkable restriction sites for Fse I, this restriction enzyme works in conjunction with the Dpn I and Eag I. b) Construct a restriction constitute of the plasmid and draw it below.Cloning StrategiesQuestion 4 (28%)Describe outline cloning strategies, including vector types (individual vectors essential not be specified) and methods used at each comprise, for the following scenariosWorked exampleYou wish to isolate the cryptogram sequence of a human coloured enzyme. You have purified the corresponding bovine enzyme and have raised a polyclonal antibody against it.- Make a cdesoxyribo nucleic acid library from human liver tissue - this will be enriched for thegenes for liver enzymes.- Create the library in an expression vector with a strong promoter so the genesare expressed in the host.- Screen the induced expression library for the presence of the desired liverenzyme utilize the bovine polyclonal antibody. The antibody will bind to thecolonies which produce the protein they recognise. Although the match maynot be exact there should be enough conserved homology to ensurerecognition.- Positive colonies will be identified by visualising the mark off on the boundantibody/secondary antibody in the colony hybridisation.a) You have a cDNA clone containing the 900 bp cryptography sequence of a cell surfaceprotein from pygmy goat monocytes. How can you use this to find thehomologous cDNA from the merino sheepb) Having...The results are as followsstep. f1 IG SEQUENCE to make single stranded DNA for sequencing UNIVERSAL PRIMER SEQUENCE for primer to anneal to, to initi ate sequencing SELECTABLE MARKER (eg lacZ) to allow selection of clones containing the chisel in MCS POLYLINKER insert fragment of DNA here 3.0 kbYou must describe the function of the essential features of each plasmid and give some indication of the plasmid size. For expression vectors you must bear in mentality the host cells in which the coding sequence will be expressed.a) Nonsense The nonsense-mediated mRNA decay way degrades mRNAs transcribed from genes in which an amino-acid codon has changed to a nonsense codon this prevents the translation of such mRNAs into truncated, and potentially harmful, proteins.c) Splicing A stage in the processing of mRNA, occurring only in eukaryotic cells, in which intervening sequences (introns) are removed from the uncreated RNA transcript (hnRNA) and the codig exons are joined together to form the mature mRNA molecule. urlwww.geneontology.org .d) booster amplifier A nucleotide sequence of DNA to which RNA polymerase binds and initiates tr anscription. It usually lies upstream of (5 to) a coding sequence. A promoter sequence aligns the RNA polymerase so that transcription will initiate at a special(prenominal) site.e) Reading Frame A series of triple