Complete 18 pages APA formatted article: Comparative Genomics. The sequencing of the positive clones was later done. The heart of the fish was used to construct cDNA pools that were later amplified using the smart cDNA library construction kit. The MYH primers were used as the probes. The PCR products were cloned, sequenced, and named depending on the organ of origin which was majorly the heart (cardiac).
In fish, the boundaries of introns and exons were identified using the Genscan. Results were verified by screening their sequence structure of the splicing of the intron based on the flanking regions of the exons. The length of the genes was estimated from the sites where the first start codon (AUG) and the third stop codon (UGA) were identified. The inter-gene distance was determined from the first nucleotide from the stop codon and the last neighboring nucleotide before the next start codon. The untranslated region, therefore, will lead to overestimation of the inter-gene distance and underestimation of the gene length. A qualitative comparison of the fish and human intron-exon organization was done.
The nucleotide sequences of our samples were analyzed using the public database from the NCBI. The sequences were biologically aligned using the BLAST program that is present in the NCBI databases too. The gene organization details were obtained from the NCBI and the Ensembl Genome Browser databases (Rossi et al., 2009).
For the muscle samples, rat samples were reared in a convectional colony under controlled conditions of 25oc, 12 hours daylight, and night each and 50% relative humidity. Water and food were provided under the veterinary regulations and guidelines. After three months, the rats were killed using the CO2 gas asphyxiation by skilled personnel. Muscles were quickly removed and stored under liquid nitrogen at a temperature of -80oc. The human muscle biopsy was obtained from the neuromuscular tissue bank with all the ethics approved (Bourque and Mabrouk, 2006). Similarly, the extraocular eye muscle samples were obtained from the Veneto eye bank foundation with all the ethics observed. They were frozen in the iso-pentane coupled with the liquid nitrogen and stored at -80oc until use (Parrington and Coward, 2007).