Each well is loaded with products from a different reaction. The strand being sequenced will have the opposite Watson-Crick base, meaning the well loaded with products of the G reaction will identify the C nucleotide in the sequence. Finally, autoradiography is performed following the electrophoretic gel separation. The polyacrylamide gel is placed in contact with a x-ray film sheet. Dark bands are produced by exposure of the 32P with the x-ray film. The sequence read from the autoradiograph will be the complementary to the single-stranded DNA template. Overall this process is also referred to as the Sanger method. The gel is read backwards from top to bottom to produce the stand in a 5’ to 3’ manner. Once the gel is read, the sequence of nucleotides is submitted to the NCBI website using BLASTN algorithm. The search options can be specified for humans, mice, or others. The option “others” was used in the case of our experiment. Once the search results are complete, the name of the gene, species, amino acid count information, among other information can be found. Results are listed in order
Each well is loaded with products from a different reaction. The strand being sequenced will have the opposite Watson-Crick base, meaning the well loaded with products of the G reaction will identify the C nucleotide in the sequence. Finally, autoradiography is performed following the electrophoretic gel separation. The polyacrylamide gel is placed in contact with a x-ray film sheet. Dark bands are produced by exposure of the 32P with the x-ray film. The sequence read from the autoradiograph will be the complementary to the single-stranded DNA template. Overall this process is also referred to as the Sanger method. The gel is read backwards from top to bottom to produce the stand in a 5’ to 3’ manner. Once the gel is read, the sequence of nucleotides is submitted to the NCBI website using BLASTN algorithm. The search options can be specified for humans, mice, or others. The option “others” was used in the case of our experiment. Once the search results are complete, the name of the gene, species, amino acid count information, among other information can be found. Results are listed in order