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Literature DB >> 8650002

Controlled ribonucleotide tailing of cDNA ends (CRTC) by terminal deoxynucleotidyl transferase: a new approach in PCR-mediated analysis of mRNA sequences.

Abstract

Controlled ribonucleotide tailing of cDNA ends (CRTC) by terminal deoxynucleotidyl transferase is a polymerase chain reaction (PCR)-mediated technique that was developed to facilitate cloning and direct sequence analysis of complete 5'-terminal unknown coding regions of rare RNA molecules. In contrast with standard tailing protocols using dNTPs as the substrate, ribo-tailing of cDNA ends is easily controllable, self-limited (from two to four rNMP incorporations) and highly efficient (>98%). By virtue of the homopolymeric ribo-tail, the modified cDNA is anchored to the 3' overhang of a double-stranded DNA-adaptor in a T4 DNA ligase-dependent ligation. PCR amplification, mediated by two sequence-specific primers, yields the desired unique product suitable for cloning and dideoxy-sequencing.

Entities: Chemical

Mesh：

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Year: 1996 PMID： 8650002 PMCID： PMC145852 DOI： 10.1093/nar/24.9.1789

Source DB: PubMed Journal: Nucleic Acids Res ISSN： 0305-1048 Impact factor: 16.971

9 in total

1. Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support.

Authors: T Hultman; S Ståhl; E Hornes; M Uhlén
Journal: Nucleic Acids Res Date: 1989-07-11 Impact factor: 16.971

2. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.

Authors: M A Frohman; M K Dush; G R Martin
Journal: Proc Natl Acad Sci U S A Date: 1988-12 Impact factor: 11.205

3. Immunoglobulin light-chain structural gene sequences cloned in a bacterial plasmid.

Authors: J G Seidman; M H Edgell; P Leder
Journal: Nature Date: 1978-02-09 Impact factor: 49.962

4. Synthetic polynucleotides. Enzymic synthesis of ribonucleotide terminated oligodeoxynucleotides and their use as primers for the enzymic synthesis of polydeoxynucleotides.

Authors: R Roychoudhury; H Kössel
Journal: Eur J Biochem Date: 1971-10-14

5. Enzymatic addition of fluorescein- or biotin-riboUTP to oligonucleotides results in primers suitable for DNA sequencing and PCR.

Authors: G L Igloi; E Schiefermayr
Journal: Biotechniques Date: 1993-09 Impact factor: 1.993

6. Terminal transferase: use of the tailing of DNA and for in vitro mutagenesis.

Authors: G Deng; R Wu
Journal: Methods Enzymol Date: 1983 Impact factor: 1.600

7. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase.

Authors: R Roychoudhury; E Jay; R Wu
Journal: Nucleic Acids Res Date: 1976-04 Impact factor: 16.971

8. Oligodeoxyribonucleotide ligation to single-stranded cDNAs: a new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification.

Authors: J B Edwards; J Delort; J Mallet
Journal: Nucleic Acids Res Date: 1991-10-11 Impact factor: 16.971

9. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency.

Authors: H Land; M Grez; H Hauser; W Lindenmaier; G Schütz
Journal: Nucleic Acids Res Date: 1981-05-25 Impact factor: 16.971

9 in total

14 in total

1. In vivo, high-resolution analysis of yeast and mammalian RNA-protein interactions, RNA structure, RNA splicing and ribozyme cleavage by use of terminal transferase-dependent PCR.

Authors: H H Chen; D Castanotto; J M LeBon; J J Rossi; A D Riggs
Journal: Nucleic Acids Res Date: 2000-04-01 Impact factor: 16.971

2. Use of terminal transferase-dependent antisense RNA amplification to determine the transcription start site of the Snrpn gene in individual neurons.

Authors: V L Buettner; J M LeBon; C Gao; A D Riggs; J Singer-Sam
Journal: Nucleic Acids Res Date: 2000-04-01 Impact factor: 16.971

Controlled ribonucleotide tailing of cDNA ends (CRTC) by terminal deoxynucleotidyl transferase: a new approach in PCR-mediated analysis of mRNA sequences.

1. Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support.

2. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer.

3. Immunoglobulin light-chain structural gene sequences cloned in a bacterial plasmid.

4. Synthetic polynucleotides. Enzymic synthesis of ribonucleotide terminated oligodeoxynucleotides and their use as primers for the enzymic synthesis of polydeoxynucleotides.

5. Enzymatic addition of fluorescein- or biotin-riboUTP to oligonucleotides results in primers suitable for DNA sequencing and PCR.

6. Terminal transferase: use of the tailing of DNA and for in vitro mutagenesis.

7. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase.

8. Oligodeoxyribonucleotide ligation to single-stranded cDNAs: a new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification.

9. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency.

1. In vivo, high-resolution analysis of yeast and mammalian RNA-protein interactions, RNA structure, RNA splicing and ribozyme cleavage by use of terminal transferase-dependent PCR.

2. Use of terminal transferase-dependent antisense RNA amplification to determine the transcription start site of the Snrpn gene in individual neurons.

3. The effect of chromatin structure on cisplatin damage in intact human cells.

4. Mapping dispersed repetitive loci using semi-specific PCR cloning and somatic cell hybrid mapping.

5. Localization of proteins bound to a replication origin of human DNA along the cell cycle.

6. A new rapid amplification of cDNA ends method for extremely guanine plus cytosine-rich genes.

7. Multiple isoform recovery (MIR)-PCR: a simple method for the isolation of related mRNA isoforms.

8. DNA polymerization catalysed by a group II intron RNA in vitro.

9. Genome-wide analysis of DNA replication and DNA double-strand breaks using TrAEL-seq.

10. Differential localization of nuclear-encoded tRNAs between the cytosol and mitochondrion in Leishmania tarentolae.