transcriptase Protocols

Category: RNA Protocols: 3' RACE Rapid Amplification of cDNA Ends Protocols

PCR cycle steps, protocol using SuperScript II reverse transcriptase (Life Technologies) and Taq polymerase, Gene specific primer, T17 Adapter primer and an Adapter primer. Dr.Dawson, Nottingham. - [Read 3' Rapid Amplification of cDNA Ends (RACE) Protocol]

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase) and to analyze the size of the DNA strand after digestion of DNA-RNA hybrids with nucleases such as S1. - [Read Alkaline Agarose Gel Electrophoresis Protocol]

Category: PCR Protocols: cDNA Synthesis Protocols

This method, for the selective amplification of full-length cDNA ends, involves the addition of an adapter during reverse transcription.  This method takes advantage of the propensity of Moloney murine leukemia virus reverse transcriptase (MMLV RT) to append two to four cytosines to the 3'-end of newly synthesized cDNA strands.  The additional residues are added when the enzyme reaches the 5'-cap structure at the end of the mRNA template. - [Read Cap-Switching RACE Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Category: RNA Protocols: RT-PCR and cDNA synthesis

Protocol for First-strand cDNA Synthesis (PCR template generation with M-MuLV Reverse Transcriptase) - [Read Protocol for First-strand cDNA Synthesis Fermentas]

Category: DNA Microarray Protocols

Reverse transcriptase labeling with Cy3/Cy5, Staining of total DNA on slides, construction, Laser scanning, Spot quantitation, Statistics, sample tracking. Vodkin Laboratory. U.Illinois. - [Read Protocols for microarray development in use in the Vodkin Laboratory]

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

This protocol describes a method for reverse transcriptase (RT) in situ PCR. In situ PCR differs from PCR in situ hybridization in the inclusion of a reporter molecule in the amplification step. The two steps of RT in situ PCR that differ from in situ PCR are overnight digestion in RNase-free DNase that is performed after protease digestion, and an RT step, prior to in situ PCR. - [Read Reverse Transcriptase In Situ PCR Protocol]

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Protocol describes a method for reverse transcriptase (RT) in situ PCR.  In situ PCR differs from PCR in situ hybridization in the inclusion of a reporter molecule in the amplification step.  The two steps of RT in situ PCR that differ from in situ PCR are overnight digestion in Rnase-free Dnase that is performed after protease digestion, and an RT step, prior to in situ PCR. - [Read Reverse Transcriptase In Situ PCR Protocol]

Category: DNA Protocols: Genomic DNA Library Protocols

Amplified cDNAs containing functional 3' and 5' splice sites are selected by digestion with BstXI and cloned into a Bluescript vector. - [Read Reverse Transcriptase-PCR for Exon Trapping and Amplification]

Category: RNA Protocols: RT-PCR and cDNA synthesis

Reverse Transcriptase-PCR Lazo Lab/Jim Hutchins - [Read Reverse Transcriptase-PCR Lazo Lab]

Category: DNA Protocols: cDNA Library Protocols

Protocol for the construction of construction of cDNA libraries by synthesis of first-strand cDNA catalyzed by reverse transcriptase. - [Read Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase for Construction of cDNA Libraries]