display was performed using GeneFishingTM DEG Premix Kits (Seegene) according to the instruction manual provided.
Three amplification fragments (CICR core cDNA
fragment, 3'- and 5'- flanking amplification fragment) were spliced together at overlapping regions using Vector NTI 10 (Invitrogen, USA).
The Fa2, Fb2 and Fc2 primers and two external primers, 5' forward FfgsII and 3' reverse RfgsII, were used to generate the second synthetic human IFN[alpha]2 cDNA
Probe preparation and labeling: The subtracted cDNA
was labeled by non-radioisotopic method using ECL random-prime labeling-detection system version II (Amersham Life Science, U.S.A.).
Researchers can isolate the mRNA generated in a cell type or tissue and copy it into molecules known as cDNA
, which then can be used for further analysis.
Polymerase chain reaction was performed in a 20-[micro]L volume reaction containing 2 [micro]L cDNA
template and 1 [micro]M each of RPL3F and RPL3R primers in HiQ-PCR Mix (Genotech).
The 3', 5'RACE and full length sequences were 635 bp (3ends), 620 bp (5ends), and 1,076 bp (SPLUNC1 cDNA
) respectively, each having 99% identity to Ovis.aries mRNA.
Two micrograms of total RNA was used for first strand cDNA
synthesis using a PrimeScriptTM RT Reagent Kit (TaKaRa).
For analysis of both the reference material and clinical samples, 1 [micro]L of cDNA
was added to each reaction.
As a template, randomly selected 3072 clones were used for PCR amplification of the cloned cDNA
by M13 universal primers.
that synthetically created cDNA
was not naturally occurring because cDNA
Extracting RNA and composing cDNA
: A 100 mg flower tissue of T.
In this study a full-length enriched cDNA
library was successfully constructed from Siberian tiger.
was used for CD19 gene amplification by conventional PCR method using Pfu DNA polymerase.
It did say that you can patent complementary DNA (cDNA
), which, it was argued, is not naturally occurring.