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Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3837

Category: PCR Protocols: cDNA Synthesis Protocols

An oligodeoxynucleotide primer hybridized to mRNA is extended by an RNA-dependent DNA polymerase to create a cDNA copy that can be amplified by PCR. Depending on the purpose of the experiment, the primer for first-strand cDNA synthesis can be specifically designed to hybridize to a particular target gene, or a general primer such as oligo(dT) can be used to prime cDNA synthesis from essentially all mammalian mRNAs - [Read Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol]

First strand cDNA synthesis with Ready-To-Go Beads Protocol - http://www.methods.info/Methods/cDNA/cDNA_ready_to_go.html

Category: PCR Protocols: cDNA Synthesis Protocols

This cDNA synthesis system simplifies your work dramatically. All reaction components are premixed and lyophylised. You have to add your RNA and (for Your-Prime beads) the primer. Another advantage of the system is a little number of pipetting steps required, and therefore reduced risk of Rnase contamination and RNA degradation. - [Read First strand cDNA synthesis with Ready-To-Go Beads Protocol]

How should I prime my cDNA? - http://invitrogen.custhelp.com/cgi-bin/invitrogen.cfg/php/enduser/std_adp.php?p_sid=HczfJTng&p_lva=1&p_faqid=1

Category: RNA Protocols: RT-PCR and cDNA synthesis: RT-PCR and cDNA Synthesis FAQ

How should I prime my cDNA? Invitrogen - [Read How should I prime my cDNA?]

In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3813

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for in vitro mutagenesis using double-stranded DNA templates. Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA. - [Read In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI]

Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_41-43.pdf

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, biotin, or fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotin- or Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes - http://www.roche-applied-science.com/PROD_INF/MANUALS/InSitu/pdf/ISH_41-43.pdf

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, Biotin, or Fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotinor Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

Protocol for siRNA-Primed RNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4326

Category: RNAi Technology Protocols

Short interfering RNAs (siRNAs) can be used to prime RNA synthesis by the RNA-dependent RNA polymerase (RdRP). SiRNAs can be used by RdRP as primers for specific cellular mRNAs, forming dsRNA products capable of inducing transitive RNAi. - [Read Protocol for siRNA-Primed RNA Synthesis Protocol]

Random Prime Labeling of DNA Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/R1.html

Category: Nucleic Acid Modification Protocols: Random Primer Labeling Protocols

Protocol for random prime labeling of DNA. - [Read Random Prime Labeling of DNA Protocol]

Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4061

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

A synthetic oligonucleotide annealed to single-stranded DNA derived from a recombinant bacteriophage M13 or phagemid template is used to prime the synthesis of complementary radiolabeled DNA. Synthesis is catalyzed by the Klenow fragment of E. coli DNA polymerase I, which extends the annealed primer for various distances along the single-stranded template DNA. - [Read Synthesis of Single-stranded DNA Probes of Defined Length from Bacteriophage M13 Templates Protocol]

Articles (1-4 of 4)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

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