A good quality total RNA was used for the first strand
cDNA synthesis using SuperScript III First-Strand Synthesis System for RT-PCR (Invitrogen, USA) according to the manufacturer recommendations.
3'- and 5'- RACE-Based Cloning of CIGR
cDNA Fragments
Broad antigenic coverage induced by vaccination with virus-based
cDNA libraries cures established tumors.
The human interferon [alpha]2b
cDNA was cloned as described by Rabhi-Essafi et al., 2007.
Construction of [L.sub.3]
cDNA library: The mRNA (3 [micro]g) purified by the above method was subjected to first strand synthesis driven by the random hexamer and cloned Murine Reverse Transcriptase using TimeSaver
cDNA synthesis kit (Pharmacia Biotech, USA).
cDNA MICROARRAYS HELP IDENTIFY GENES INVOLVED IN AD
Oligonucleotides and 5 x HiQ-PCR mix were obtained from Genotech (Daejeon, Korea) and M-MLV
cDNA synthesis kits were purchased from Enzynomics (Daejeon, Korea).
Synthesis of
cDNA was followed by standardization of PCR and finally content of IRS-2 was detected by PCR.
The 3ends were synthesized according to the instructions in 3'Full RACE Core Set
cDNA Kit (Takara Biotechnology, China).
For
cDNA cloning and tissue expression analysis, three individuals with an average body weight of 11.9+-0.62g were deeply narcotized with an overdose of tricaine methanesulfonate (MS-222) (Sigma) after 24-hour food deprivation.
Archived
cDNA material from 70 CML clinical samples encompassing 7 different disease levels (as measured by RT-qPCR and expressed as a percentage of [BCR-ABL1.sup.IS]) were used: 20%, 10%, 1%, 0.1%, 0.01%, 0.001%, and <0.001% BCR-ABL1 (see online Supplemental Table S2).
First of all, mammary tissues of Kivircik sheep were collected in two stages, before parturition and after parturition (the secretion of colostrums known to be considerably high), and two different
cDNA libraries were constructed from those tissues.