PCL366H1 Lecture Notes - Lecture 1: Hemocytometer, Lysis Buffer, Ribosomal Rna
Document Summary
Small interfering rna (dsrna molecules) of 20-25 bp that interfere expression of specific genes by forming complementary nucleotide sequences, leading to breakdown of mrna. Process of introducing nucleic acids into cells. Done through viruses or opening pores/holes in the cell membrane to allow uptake of materials (in this lab, dharmafect was used with the pore method) rrna. Consists of 28s (larger piece used to control gene expression and indicate good quality) and 18s (smaller piece, same as 28s) Measures 260 nm and 280 nm wavelengths and tells of the purity of rna (1. 7 1. 9 is. Tests for successful formation of pcr products (graphs at the end of rt-pcr). High success rate will be indicated by 1 large uniform peak. Secondary peaks imply formation of primer-dimers or aborted, short pieces. Making cdna from rna using reverse transcriptase. Pcr: denature (94 - 95 c) , anneal (50-60 c), extend (72 c)
