Advantage RT-for-PCR Kit

Description

Complete kit for the synthesis of first-strand cDNA from total RNA or polyA+RNA. The kit contains sufficient reagents for 100 first-strand cDNA synthesis reactions. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back The reliability of cDNA synthesized with the Advantage RT-for-PCR Kit.The time course of lipopolysaccharide induction of iNOS mRNA in the mouse macrophage cell line RAW 264.7 was studied by isolating RNA from culture samples taken at the time points indicated in hr . Approx. 0.4 mug was then reverse transcribed using the Advantage RT-for-PCR Kit, and 5 % of the cDNA product was used as a template for PCR using the Mouse G3PDH Amplimer Set Cat. # 639009 as a control Panel A and mouse iNOS primers Panel B . 20 % portions of the PCR products were run on a 1.8 % EtBr/agarose gel. Lane M: X174/HaeIII DNA size marker. Back PCR amplification of cDNA synthesized with the Advantage RT-for-PCR Kit vs. cDNA synthesized with kits from two competitors.The indicated amounts of Human Lung Total RNA Cat. # 636507 were used for reverse transcription, and 5 % of each cDNA product was amplified by PCR using the Human G3PDH Control Amplimer Set Cat. # 639005 . Lanes 1, 5 & 9: 250 ng. Lanes 2, 6 & 10: 62.5 ng. Lanes 3, 7 & 11: 16 ng. Lanes 4, 8 & 12: 4 ng. Lane M: X174/Hae III DNA size marker. The results show that when 4 ng of RNA was used as the starting material, only the Advantage RT-for-PCR Kit generated a clearly visible product. Back