Using a kinetic approach, a cDNA library composed of almost equal representations of all genes expressed in the aerial parts of 2โweekโold __Arabidopsis__ was constructed. A cDNA was synthesized with an oligo dT primer containing a __Not__l site. A linker containing the nucleotide sequence of __Sse_
Construction of cDNA libraries from cultures of Armillaria mellea
โ Scribed by Joe O'Connell; Helen M. Sheehan; Shawn Doonan
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 875 KB
- Volume
- 25
- Category
- Article
- ISSN
- 0020-711X
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A simplified cDNA synthesis and cloning method, suitable for efficient generation of cDNA libraries at frequencies up to lo6 clones/pg mRNA, is described. Routine synthesis oftranscripts of well over 4 kb is facilitated by the use of high-quality RNA template isolated from materials rich in RNases.
We describe here a general and simple procedure for cDNA library construction making use of in vitro amplification of cDNA by polymerase chain reaction (PCR). The first-strand cDNA is synthesized from total RNA with a primer EcoRI-(dT)17 and oligo(dG) tailed. An oligonucleotide, EcoRI-BamHI-(dC)13,