Aptamer-dependent full-length cDNA synthesis by overlap extension PCR
نویسندگان
چکیده
منابع مشابه
Large-scale RT-PCR recovery of full-length cDNA clones.
Pseudogenes, alternative transcripts, noncoding RNA, and polymorphisms each add extensive complexity to the mammalian transcriptome and confound estimation of the total number of genes. Despite advanced algorithms for gene prediction and several large-scale efforts to obtain cDNA clones for all human open reading frames (ORFs), no single collection is complete. To enhance this effort, we have d...
متن کاملFull-length cDNA synthesis for long-distance RT-PCR of large mRNA transcripts.
Reverse transcription of mRNA often leads to the synthesis of partial, non-full-length cDNAs. Methods to facilitate reverse transcription across RNA regions of secondary structure, as well as enzyme modifications to eliminate RNase H activities inherent to reverse transcriptase enzymes, have been previously reported. However, because all reverse transcriptases have high error rates of polymeriz...
متن کاملCloning of full-length cDNA of teleost corticotropin-releasing hormone precursor by improved inverse PCR.
We designed a new inverse PCR protocol combined with switching mechanism at 5' end of RNA transcript (SMART) technology, and applied it to the cloning of teleost corticotropin-releasing hormone precursor cDNA. Due to the advantages of both techniques, this method can efficiently amplify the complete 5'- and 3'-ends of cDNA in a single reaction, and might prove to be an alternative to the conven...
متن کاملA Nested-Splicing by Overlap Extension PCR Improves Specificity of this Standard Method
Background: Splicing by overlap extension (SOE) PCR is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function. Objectives: We introduced a nested-SOE-PCR (N –SOE-PCR) in order to increase the specificity and generating mutations in a gene by SOE-PCR. Materials and Methods: Genomic DNA from Bacillus thermo...
متن کاملAn efficient full-length cDNA amplification strategy based on bioinformatics technology and multiplexed PCR methods
A novel strategy for amplification full-length cDNA and promoter sequences has been developed using bioinformatics technology and multiplexed PCR methods in this study. The amplification of 3' ends of cDNA is performed according to the modified classic 3' RACE techniques, therein the more efficient and effective oligo(dT)-anchor primer with hairpin structure is specially designed. For the ampli...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioTechniques
سال: 2004
ISSN: 0736-6205,1940-9818
DOI: 10.2144/04371dd02