Notes: Using the T7 RiboMAX™ Express Large Scale RNA Production System RNA standards were created for a highly sensitive real-time qPCR method. (5191)

Notes: The T7 RiboMAX™ Express Large Scale RNA Production System RNA was used to amplify desired sequences for use in electrophoretic mobility-shift assay (also known as gel-shift, mobility shift, or EMSA) to study RNA:protein interactions. (5192)

Notes: This paper reports on the use of the T7 RiboMAX™ Express Large Scale RNA Production System to perform in vitro transcription, for the study of viral RNA structures and their functional relevance. (5190)

Notes: The T7 RiboMAX™ Express Large Scale RNA Production System was used for the transcription of RNA for viral inoculation through electroporation. (5189)

Notes: This paper reports on the use of the T7 RiboMAX™ Express Large Scale RNA Production System for the generation of dsRNA for RNAi, to determine function(s) of a gene critical for viral infection. (5188)

Notes: These authors used the Flexi^® Vector System to prepare ORF clones encoding 1929 human genes and to transfer a subset of these clones to various expression vectors for further analysis. They created HaloTag^® fusion proteins and examined expression of these proteins in vitro and in COS7 and HEK293 cells. They also performed comparisons between the Flexi^® System and Gateway^® cloning system, specifically examining the effects of flanking sequences on protein expression in in vitro translation systems and confirming that the cellular localization of the HaloTag^® fusion proteins was consistent with results obtained using GFP-fusions. (3800)

Notes: To identify microRNA targets, the authors created a Drosha-knockdown cell line and confirmed depletion of Drosha and three randomly selected miRNAs in these cells by quantitative RT-PCR, using β-actin as a control. The reverse transcription step was performed using the ImProm-II™ Reverse Transcription System. The authors then performed microarray analysis to monitor expression of transcripts to determine which were upregulated as a result of Drosha depletion; cRNA used in these microarray experiments was synthesized using the T7 RiboMAX™ Express Large Scale RNA Production System. Cyclin D1 was identified as a potential miRNA target. To screen miRNAs that regulate cyclin D1, the authors cloned the cyclin D1 3´ untranslated region downstream of the firefly luciferase gene of the pGL3-Control Vector and measured luciferase levels in transfected cells using the Dual Luciferase Reporter Assay System. Renilla luciferase in the pRL-TK Vector was used as a normalization control. (3894)

Notes: The authors developed a method to reverse transcribe poly(A)+ RNA from leukocytes without using oligo(dT) immobilized on a 96-well plate. Four RNA targets, as well as a synthetic control RNA, were reverse transcribed using MMLV Reverse Transcriptase (1X reverse transcription buffer [50mM KCl, 10mM Tris-HCl (pH 8.3), 5.5mM MgCl₂, 1nL/µL Tween 20], 1.25 mM each dNTP and 4 units of Recombinant RNasin^® Ribonuclease Inhibitor), then quantitated by real-time quantitative PCR. The authors varied the concentration of MMLV Reverse Transcriptase, incubation time, and primer/template ratio] to obtain the maximum yield. Small quantities of MMLV Reverse Transcriptase were sufficient to reverse transcribe short synthetic RNA and abundant RNAs, but approximately 100 units was required for other RNAs. The synthetic control RNA was synthesized using the T7 RiboMAX™ Express Large Scale RNA Production System. (3456)