Objective: To develop Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) method for the estimation of adenosine A 1 receptor mRNA levels in rat kidney. Material and Methods: Total cellular RNA was isolated from whole rat kidney by small-scale total RNA preparation protocol and it was reverse transcribed into cDNA. The cDNA was subjected to PCR amplification using gen specific primers. The...

Full-length cDNAs play an essential role in identifying genes and determining their promoter regions. Here we describe a simple method for constructing a full-length cDNA library, which has the following advantages: (i) it consists of only three steps including direct ligation between a vector and a cDNA strand using T4 RNA ligase, (ii) it contains neither a PCR process generating mutations nor...

Cloned cDNAs coding for hepatic UDP-glucuronyltransferase (UDPGT) have been isolated from a rat liver cDNA library in the expression vector bacteriophage lambda gt11 using anti-UDPGT antibodies. Four different mRNAs have been identified by sequencing of 15 UDPGT cDNA clones. The sequences of the four classes of cDNA were determined to be 85-95% homologous. Restriction fragments were isolated fr...

Complementary DNA (cDNA) specific for gamma-globin nucleotide sequences has been prepared by hybridizing total cDNA made from cord blood messenger RNA (mRNA) as template to an excess of normal adult human globin mRNA and recovering the single-stranded cDNA from hydroxylapatite. The specificity of the gamma cDNA for gamma mRNA sequences is strongly supported by the hybridization of this cDNA at ...

Eight complementary DNA (cDNA) clones highly expressed in fetal rat stomach but not in normal adult rat stomach were isolated after screening 2 x 10(4) independent recombinants from a subtracted cDNA library. The cDNA library was first prepared from RNAs of total stomach at 16 days gestational period, and this cDNA library was subtracted by cDNAs prepared from adult rat total stomach RNA, using...

human growth hormone (hgh) genomic sequence containing 5 exons and 4 introns was cloned in pcdna-3 and the constructed plasmid was subsequently used for transfection ofnlli-3t3 cell line using lipofection technique. expression of hgh in stably transfected cells was assayed using elisa. total rna was extracted from transfected cells and hgh cdna was amplified by rt-pcr using specific primers. th...

background: a major issue in many gene expressionstudies utilizing small amount of biological materials is thelimited quantity of rnapurified from clinical samples, which isoften used for rt-pcr or standard northern blot analysis.objectives: the smart cdna synthesis method and subsequentsmart-cdna-pcr technique was used to analyse 3genes in macroschizonts of theileria annulata in small lymphnod...