cDNA Synthesis Kits - SMART cDNA Synthesis - Clontech
Clontech's SMART cDNA Synthesis Kit - Biocompare
In an eukaryotic cell, the mRNA population constitutes approximately 1% of total RNA with the number of transcripts varying from several thousand to several tens of thousands. Normally, the high abundance transcripts (several thousand mRNA copies per cell) of as few as 5-10 genes account for 20% of the cellular mRNA. The intermediate abundance transcripts (several hundred copies per cell) of 500-2000 genes constitute about 40-60% of the cellular mRNA. The remaining 20-40% of mRNA is represented by rare transcripts (from one to several dozen mRNA copies per cell) [Alberts , 1994]. Such an enormous difference in abundance complicates large-scale transcriptome analysis, which results in recurrent sequencing of more abundant cDNAs.
A review of the Clontech's SMART cDNA Synthesis Kit
Transcriptional profiling using the common microarray platforms, whether in-house two colour slide based, Agilent, Affymetrix or Illumina all require microgram quantities of RNA and as such impose severe restrictionson the type of biological samples that can be used bystandard non-amplified protocols. However, with an ever-increasing need to utilise smaller andsmaller samples and address the relevant questions in the clinic, techniques have beendeveloped to amplify whole RNA populations from material such as clinicalbiopsy or laser capture microdisection and others. These techniques are now routinelyused by many commercial venders. Some of the more common approaches arereviewed here.
cDNA synthesis/reverse transcription kit review - Labome
One of the greatest advantages of SMART technology is its increased efficiency compared to traditional technologies such as adaptor ligation. Its high efficiency and sensitivity enables you to use a very limited quantity of starting material, such as microdissected tissues, laser-captured cells, biopsy samples, etc. As little as 1-2 ng of total RNA is sufficient for generating a highly representative cDNA pool for different downstream applications.
SMART PCR cDNA SYNTHESIS Kit ..
With this experimental design, 130 plasmid clones were obtained. The digested cDNA clones were searched for sequence homologies in the GenBank DNA database by BLAST. DNA sequencing results of the digested 130 clones were found to have high homology to 14 genes known in the public database of the 14 kinds (). We identified 21 unknown genes, which were not homologous to any of the known genes in the Genbank database (). Among these 21 genes, we selected 7 genes which were over- expressed in glioblastoma, compared to non-tumor brain tissue (). To be considered over-expressed in glioblastomas, the genes tested had to show a threefold greater expression in tumor samples than in non-tumor brain tissue by semiquantitative RT-PCR. In this study, we considered these genes as candidates for over-expressed genes in glioblastoma.