Macvector recombination software#
Data were collected, assembled, and edited on a PowerMac using inherit software (Applied Biosystems). Therefore, we obtained this clone from the IMAGE consortium at Lawrence Livermore National Laboratory and it was sequenced by the Sanger dideoxynucleotide chain-termination procedure in a stepwise fashion by primer walking. 153195) that is a cDNA sequence derived from human breast tissue cloned into pT3T7 vector (Pharmacia) and bearing significant homology to REC2 ( P = 0.90). This sequence and the Rec2 sequence containing the RecA core were used to query the database of expressed sequence tags by the blastx program. One clone amplified from human kidney cDNA with GKTQ-f and STIY-r produced a 110-bp fragment that contained significant homology to Rec2. Approximately 60 gene fragments were cloned into the pCRII plasmid (Invitrogen) by T-A cloning and sequenced with a fluorescent PCR-based Sanger dideoxynucleotide chain-termination procedure. These primers were used in the PCR using the Expand system (Boehringer), and after two rounds of amplification, the bands were purified on a 3% SeaPlaque (FMC) low-melting-temperature agarose TAE gel. This was accomplished by using human codon usage tables and incorporating the inosine nucleotide into DNA primers on an Applied Biosystems DNA synthesizer. maydis REC2 gene, a set of 10 primers were designed with codon degeneracy. Based on sequences in the conserved and divergent regions of the U. Degenerate primers were used for PCR amplification.
0 kommentar(er)
