Notes: These authors used Promega's AMV Reverse Transcriptase, RNasin^® Ribonuclease Inhibitor and Taq DNA Polymerase in this study. (2061)

Notes: Promega's Access RT-PCR System was used in this study. (1997)

Notes: PCR was performed using primers to introduce a T7 promoter and a translation initiation sequence into PCR products used as templates in TNT^® Coupled Reticulocyte Lysate System reactions. Truncation mutations were detected in 12/26 FAP patients; however, 33% of confirmed protein truncations in APC were not detected using the protein truncation test (PTT). (1238)

Notes: PCR products were cloned into the pGEM^®-T Vector System. (0082)

Notes: The combination of RT-PCR and PTT were used to detect novel mutations in 4/6 DMD patients. The PTT analysis was performed using the TNT^® T7 Coupled Reticulocyte Lysate System. (0243)

Notes: The system was used to clone the amplimers generated by RT-PCR. Sequencing of the clones was performed in the vector. (1569)

Notes: C-terminal and N-terminal GT-2 deletion mutant proteins as well as site-directed GT-2 mutant proteins were synthesized from PCR templates using the TNT^® T7 Coupled Reticulocyte Lysate System. Polypeptide segments in the N-terminal and C-terminal domains of GT-2 were analyzed for differential DNA binding to GT-box target sites. (0609)

Notes: SSCP and heteroduplex analysis were used to screen exons of APC from gastric cancer lines; in addition, protein truncation test (PTT) was used to screen the MCR (mutation cluster region) of exon 15. The template was produced by PCR of genomic MCR DNA and used in the TNT^® Coupled Reticulocyte Lysate System in the presence of ³⁵S methionine. (0309)

Notes: The RNAgents^® Total RNA Isolation System was used to isolate total RNA from rat brains or 10-13 day-old primary cultures of rat cardiac myocytes. The isolated RNA was used for RT-PCR. (1657)

Notes: The pGEM^®-T Vector System was used to clone PCR products amplified from a tobacco cDNA library. (1968)

Notes: The researchers optimized the amount of PCR template for use in the TNT® T7 Coupled Reticulocyte Lysate System. They suggest a titration over a range between 0.2 to 7.5pmol/25µl reaction for each template used. (1910)

Notes: This group evaluated RNA-based screening methods for detection of mutations in hMLH1 and hMLH2. Their results suggest that PTT is useful for preliminary screening tests to localize a mutation, which then can be confirmed by sequencing. The PTT was performed using T7 polymerase and the TNT^® T7 Coupled Reticulocyte Lysate System. (0875)

Notes: The protein truncation tese (PTT) was used to analyze BRCA1 exon 11, which includes ~61% of the coding region of the gene. The observed frequencies of mutations in the BRCA1 gene were not significantly different from expected, based upon phenotype and family history. PTT was performed using the TNT^® T7 Coupled Reticulocyte Lysate System. Each mutation was confirmed by an independent PCR amplification and PTT analysis. (1258)

Notes: RT-PCR and the Protein Truncation Test (PTT) were used to identify DMD/BMD-associated point mutations that were not detected by DNA-based methods. The RNA method is more sensitive because many of the primers used for methods such as multiplex PCR do not permit the detection of mutations affecting splice sites. The RNA method used in this study screened the total coding region. PTT was performed with the TNT^® Coupled Reticulocyte Lysate System. (0491)

Notes: These researchers cloned the FHIT gene by PCR and expressed it in the TNT® Coupled Reticulocyte Lysate System. Synthesized proteins were analyzed using SDS-PAGE and autoradiography. (0596)

Notes: Taq DNA Polymerase was used in this study. (1961)

Notes: RT-PCR and PTT detected truncated peptides produced from the mRNA transcripts of mutant CFTR genes. Due to the large size of the gene for CFTR, RT-PCR and the PTT are attractive tests for screening for frameshift mutations in the gene. The PTT was performed using an aliquot of nested PCR product (consisting of the CFTR cDNA fused to the T7-promoter) in the TNT® T7 Coupled Reticulocyte Lysate System. (1869)

Notes: The fmol^® DNA Cycle Sequencing System was used to determined the sequence of cloned PCR products. (1140)

Notes: Taq DNA Polymerase was used in this study. (2013)

Notes: The gene for 20.5K protein of Ad3/7 was amplified by PCR and used as a template for the TNT® Coupled Reticulocyte Lysate System using [35S]cysteine. The proteins were immunoprecipitated with antisera to a 20.5K-specific synthetic peptide. (1920)

Notes: [³⁵S]methionine-labeled ApCREB2 and ApC/EBP were translated in the TNT^® Coupled Reticulocyte Lysate System. In vitro interaction assays were performed with the labeled ApCREB2 and ApC/EBP and with GST fusions of ApCREB2 and its deletion mutants. ApCREB2 forms weak homodimers, and ApCREB2 interacts with ApC/EBP. (1781)

Notes: The Altered Sites^® II in vitro Mutagenesis System was used to produce 14 consecutive 4bp deletions in a putative response element. (1157)

Notes: RBP-Jk protein was transcribed and translated in vitro using the TNT^® Coupled Reticulocyte Lysate System. The protein was labeled with [³⁵S] and used in mobility shifts with PCR-generated probes to demonstrate the binding of the RBP-Jk protein to the RBP-Jk site in the Cp promoter. (1800)

Notes: Promega's Taq DNA Polymerase was used in this study. (1957)

Notes: This report details the synthesis, characterization and application of a photocleavable biotin derivative (PCB) for use in detection of biomolecules, without the limitations of irreversibility (also see patent, below). The reagent, PCR-NHS contains a biotin moiety linked through a spacer arm to a photocleavable moiety that reacts selectively with amino groups on a target molecule to form a stable carbamate. The conjugate undergoes a photoreaction at 300nm, with rapid release of the unmodified target molecule. (1944)