Notes: Poly A+ RNA was isolated from mouse female fibroblast total RNA and male ES cell total RNA using the PolyATtract^® mRNA Isolation System. The isolated RNA was used for RT-PCR and RNase protection assays. (1682)

Notes: The RNAgents^® Total RNA Isolation System was used to isolate total RNA from human brain superior frontal gyrus. The RNA was used in RT-PCR. (1517)

Notes: Taq DNA Polymerase was used in this study. (2189)

Notes: Real-time quantitative RT-PCR was performed using the Access RT-PCR System. (1102)

Notes: Promega's Tth DNA Polymerase, pGEM^®-T Vector System and Riboprobe^® System were used in this study. (2002)

Notes: Eighteen sporadic neoplasms of ampullary origin were examined for the involvement of mutations in APC, Ras, p53 and chromosomal 5q21 allelic losses. 5q21 loss (including APC) occurred in 50% and APC mutations occurred in 17% of the ampullary tumors. 5q21 loss and mutations in APC and Ras occur in early tumor development, and p53 inactivation is associated with progression of malignancy. The researchers used PTT for APC mutation detection in the TNT® T7 Coupled Reticulocyte Lysate System with chemiluminescent labeling instead of radiolabeling of proteins. (2056)

Notes: In this study a variety of techniques were employed to screen the causative APC mutations in classical FAP cases. A combination of PTT and chemical cleavage of mismatch analysis was most effective. RT-PCR was used to amplify exons 1-14 from a lymphoblastoid cell line. 9 out of ten FAP kindreds had APC mutations that were detected with the chemical cleavage technique. PTT verified the results from the chemical cleavage analysis. The other mutation is linked to the APC gene and may be in the promoter region of the gene. (1848)

Notes: PTT was used to rapidly screen the BRCA1 gene. Transcription and translation was performed on five overlapping fragments (amplified by PCR) of the entire coding region of the BRCA1 cDNA or genomic DNA using the TNT^® T7 Coupled Reticulocyte Lysate System. (1144)

Notes: The protein truncation test was performed on PCR products from the BRCA2 gene. The PCR products were used in the TNT^® T7 Coupled Reticulocyte Lysate System to express the protein, and the protein was analyzed by SDS-PAGE. (0834)

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)