Second Strand cDNA
synthesis and Differential cDNA
Based on CICR cDNA
amplification fragment information, two specific primers (CIGR-GSP1 (5'-AGG GAG ATC GTG AAC CTC GTC-3'), CICR-GSP2 (5'-TTC ACC ATA GCC TCC TAC AGT CCA ACC AAA-3')) were designed for amplification of 3' and 5' end of CICR amplification fragment, respectively.
pastoris codon-optimized human interferon [alpha]2 cDNA
clones (WS, SW and FS)
Preparation of subtracted cDNA
: To identify infective stage ([L.sub.3]) specific genes, the use of subtracted probe development technique was followed.
To this end, researchers at the Mount Sinai School of Medicine in New York are studying cDNA
expression patterns in patients with known mental status-no dementia, questionable dementia, mild dementia, or moderate dementia (which corresponds to early or mild AD)-for whom brain samples are available after death.
The concentration of total RNA was adjusted to 200 ng/[micro]L for cDNA
Briefly, recombinant pMD-18T with SPLUNC1 ORF was digested with SnaB I and Not I and the SPLUNC1 cDNA
fragment was inserted into pPIC9K vector previously linearized with similar enzymes to obtain the pPIC9K/SPLUNC1 cDNA
The quantification and purity of RNA was measured by spectrophotometer (Thermo Scientific, Waltham, MA, USA).The samples with an absorption ratio (260 nm / 280 nm) between 1.8 and 2.1 were used for cDNA
samples were stored at -80 [degrees]C at the Hammersmith Hospital (for a median of 2 years, range 1-7 years) following the processing of peripheral blood samples as they arrived at the laboratory according to a previously described protocol (15-16).
Library Construction, Quality Controls, and Sequencing.
complimentary DNA (cDNA
) were patent eligible subject matter under
(2010) reported a cDNA
sequence of NTMS1 (EU375459) that controls endosperm development was cloned from Nicotiana tabacum.