Notes: The ImProm-II™ Reverse Transcriptase was used to amplify the protein coding region of a 2,264 base mouse MT-1 gene mRNA and GAPDH from mouse total RNA.  RT-PCR was accomplished with Pfu DNA polymerase.  The RT-PCR product was used to make a probe for Northern analysis. (2609)

Notes: In this paper, ImProm-II™ Reverse Transcriptase was used to make cDNAs of MusTRD using template RNA isolated from C2C12 myotubes (C2C12) and various mouse hind limb muscle tissues. One microliter of the cDNA products from the reaction was used in real-time PCR analysis using primers for the alternately spliced MusTRD products and SYBR^® Green Dye.  Data is presented as relative expression compared with the C2C12 cell line or as gel images of the resultant amplimers.  (2844)

Notes: RT-PCR was used to assess the response of mitotic spermatogonia to Kit1. Spermatogonia were cultured with and without Kit1 and RNA harvested after 24 hours. RT-PCR was performed in a two-step reaction using first the ImProm-II™ Reverse Transcriptase in a 20µl reaction. One microliter of the RT reaction was removed and amplified in a 50µl PCR using the PCR Master Mix. (2626)

Notes: Improm-II™ Reverse Transcriptase was used to make cDNAs of mouse ZIP4, a zinc metal ion transporter. The authors used 1μg of total RNA for the reverse transcription reaction. Some reverse  transcription reaction products were used to make probes for Northern blot analysis of expression in various tissues. Others were used to make mammalian expression constructs for transient transfection experiments. Details of each application are provided. (2722)

Notes: The Y-box-binding protein YB-1 was examined through the use of a reporter plasmid with wildtype and mutant putative Y-box binding sites. The binding sites were constructed in the pGL3 Basic Vector and measured using the Luciferase Assay System. Studies were performed in normal human dermal fibroblasts.  Expression of YB-1 was found to repress expression of the COL1A2 gene at the transcriptional level. Confirmation of this dose-dependent inhibition was through measurement of the steady-state mRNA levels by quantitative, real-time RT-PCR.  The reverse transcription portion of the quantitative, real-time RT-PCR reactions were performed with ImProm-II™ Reverse Transcriptase followed by a TaqMan-type quantitative PCR step. (2623)

Notes: RT-PCR was used to confirm the expression of the lef-12 gene at various times post-infection. The RT step was performed using ImProm-II™ Reverse Transcriptase in a 20µl reaction volume. One microliter of the RT reaction was amplified in a 50µl PCR amplification reaction using the PCR Master Mix. (2628)