Top : Molecular Biology : RNA : cDNA

Summary: cDNA stands for complementary DNA or copy DNA and can be single-stranded or double-stranded. cDNA is synthesized in vitro from a mRNA template using reverse transcriptase. The resulted cDNA is single-stranded. This process is called reverse transcription (RT) or first strand cDNA synthesis. In some experiments such as microarray, double-stranded cDNA is required and can be produced by another round of DNA synthesis following the first strand synthesis. The purpose of converting mRNA to cDNA is mainly for the analysis of the template mRNA because DNA is much stable than RNA. Once mRNA is converted to cDNA, the cDNA can be used for RT-PCR, as probe for expression analysis and for cloning of the mRNA sequence.
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Protocols
  • cDNA Production and Cloning
    Produce cDNA and subsequent clone it into vector. Protocol includes: 1st Strand cDNA Synthesis Determine the efficiency of first strand cDNA synthesis Second Strand cDNA Synthesis Cleave 5' Overhanging Ends Methylate to Protect EcoRI Sites in cDNA Ligate to Linkers Ligate cDNA to Vector
    http://www.omrf.org/OMRF/Research/09/FrankLab/CDNA...
    Added: Wed Feb 04 2009, Hits: 11197, Reviews: 0  Write review   Cached
  • cDNA Library Construction (Michael Blaber's Lab)
    Very useful guide and protocol for cDNA library construction
    http://wine1.sb.fsu.edu/bch5425/lect27/lect27.htm
    Added: Tue May 14 2002, Hits: 593, Reviews: 0  Write review   Cached
  • cDNA Synthesis (Cungui Mao)
    http://www.musc.edu/bcmb/ceramide/protocols/0043.h...
    Added: Tue May 14 2002, Hits: 8990, Reviews: 0  Write review   Cached
  • cDNA Synthesis (Promega)
    Provides detailed procedures for construction of cDNA library, including first strand and double-stranded cDNA synthesis from mRNA, and subsequent ligation into a suitable vector.
    http://www.promega.com/tbs/tm038/Tm038.html
    Added: Tue May 14 2002, Hits: 9686, Reviews: 0  Write review   Cached
  • cDNA/AFLP (CWB Bachem)
    The protocol which follows gives a step-by-step description of cDNA-AFLP, a method for visualisation of differential gene expression
    http://www.dpw.wau.nl/pv/staff/aflp.htm
    Added: Tue May 14 2002, Hits: 184, Reviews: 0  Write review   Cached
  • Differential cDNA Screening (Gerard Lazo)
    cDNA library construction and differential screening
    http://wheat.pw.usda.gov/~lazo/methods/misc/Talbot...
    Added: Tue May 14 2002, Hits: 660, Reviews: 0  Write review   Cached
  • ESTs from Phage Plaques (the WHO/UNDP/World Bank Schistosoma Genome Network)
    This protocol uses PCR to amplify phage insert for sequencing and EST production.
    http://www.nhm.ac.uk/hosted_sites/schisto/protocol...
    Added: Tue May 14 2002, Hits: 303, Reviews: 0  Write review   Cached
  • ESTs from Phage Plaques (the WHO/UNDP/World Bank (WHO)
    Schistosoma Genome Network)
    A slightly different protocol from the above one.
    http://www.nhm.ac.uk/hosted_sites/schisto/protocol...
    Added: Tue May 14 2002, Hits: 218, Reviews: 0  Write review   Cached
  • Library cDNA Synthesis (Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
    Dealing with first strand and second cDNA synthesis for library construction.
    http://www.nottingham.ac.uk/~mbzspd/methods/Librar...
    Added: Sat Apr 10 2004, Hits: 154, Reviews: 0  Write review   Cached
  • Protocol for First-strand cDNA Synthesis (Fermentas)
    http://www.fermentas.com/TechInfo/ModifyingEnzymes...
    Added: Tue May 14 2002, Hits: 1541, Reviews: 0  Write review   Cached
  • Screening cDNA Clones by RNA In Situ Hybridization (Berkeley Drosophila Genome Project)
    The RNA in situ procedure described below is based on the protocol developed by Tautz and Pfeifle, but adapted to allow the screening of 96 RNA probes on whole-mount Drosophila embryos at the same time.
    http://www.fruitfly.org/about/methods/RNAinsitu.ht...
    Added: Tue May 14 2002, Hits: 1009, Reviews: 0  Write review   Cached