By Hiroyuki Takahara, Elmar Endl, Richard O’Connell (auth.), Chaofu Lu, John Browse, James G. Wallis (eds.)
The quite a few important purposes of complementary DNA (cDNA) expertise have replaced dramatically because the know-how has complex over fresh years. In cDNA Libraries: tools and Protocols, professional researchers offer present ideas that replicate the newest advances within the building and alertness of cDNA libraries. the 1st half the amount covers better techniques to a couple of the main easy components of making cDNA libraries, whereas the second one part casts a wider internet and contains visionary functions of cDNA know-how which have been both unexpected or technically impractical till lately. Written within the hugely profitable Methods in Molecular Biology™ sequence structure, chapters comprise introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and key tips about troubleshooting and keeping off recognized pitfalls.
Authoritative and state-of-the-art, cDNA Libraries: tools and Protocols serves as an incredible advisor to all scientists trying to increase this significant know-how and supply solutions to the iconic primary questions of biology.
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Additional resources for cDNA Libraries: Methods and Applications
Nucleic Acids Res. 29, e29. , Modi, C. and Winegarden, N. (2002) 35 Representation is faithfully preserved in global cDNA amplified exponentially from sub-picogram quantities of mRNA. Nat. Biotechnol. 20, 940–943. , Okuzaki, D. and Nojima, H. (2008) Chum-RNA allows preparation of a high-quality cDNA library from a single-cell quantity of mRNA without PCR amplification. Nucleic Acids Res. 36, e92. Chapter 3 Construction of a Full-Length cDNA Library from Castor Endosperm for High-Throughput Functional Screening Chaofu Lu, James G.
0), 80 mM MgCl2, 20 mM spermidine, and 50 mM DTT. 0) and 10 mM MgCl2. 0), and 10 mM ethylenediaminetetraacetic acid (EDTA). Centrifuge column: CHROMA SPIN-400 (Clontech, Palo Alto, CA). Vector DNA: pAP3neo (TaKaRa Bio). 5) and 1 mM EDTA (see Note 4). 1 mM EDTA (see Note 4). Centrifuge filter: Ultrafree-C3 (Cat. #UFCP3TK50; Millipore, Bedford, MA) or MINICENT-30 (Cat. #08627; Tosoh SMD, Grove City, OH) (see Note 5). QuickPrep Micro mRNA purification (QMP) kit: Cat. , Piscataway, NJ). Plasmid Maxi kit: Cat.
2004) Optimized RNA amplification using T7-RNApolymerase based in vitro transcription. Anal. Biochem. 334, 164–174. R. and Saitou, M. (2006) An improved single-cell cDNA amplification method for efficient high-density oligonucleotide microarray analysis. Nucleic Acids Res. 34, e42. , Ohinata, Y. and Saitou, M. (2007) Global single-cell cDNA amplification to provide a template for representative high-density oligonucleotide microarray analysis. Nat. Protoc. 2, 739–752. , Yabuta, Y. and Kurimoto, K.