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The PCR product was cloned to a BglII site upstream of the flippase recognition target (FRT) in pKD13 (20).
To facilitate cloning into the vector pCAMBIA1301, a restriction enzyme site of PstI was incorporated into a forward primer, whereas a BglII site was flanked with a reverse primer.
Twenty-one alleles of the IL6 promoter (690 bps, chr7:22732707-22733396) that contained 9 rs1800797 A alleles and 12 rs1800797 G alleles were successfully amplified using primers listed in Supplemental Material, Table S2, from 13 human BEC cultures that were heterozygous for rs1800797 and were directionally cloned into the pGL2-basic luciferase reporter vector (Promega) upstream of the luciferase coding sequence using MluI and BglII cloning sites according to the manufacturer's instructions.
The PCR products were ligated into pUCm-T vector (Beyotime, China) first, and then subcloned into pEGFP-N1 with BglII and KpnI.
The amplified fragment was cloned into the NheI and BglII digested pIRES2-ZsGreen vector for overexpression of WIF-1.
Enzymatic properties and intracellular localization of the novel Trichoderma reesei p-Glucosidase BGLII (Ce11A).
33) The full-length, wild-type human XIICC'4 cDNA was obtained by polymerase chain reaction (PCR) from the cDNA pool of human T cell leukemia MOLT-4 cells and inserted into p3xFLAG-CMV10 vector using EcoRI and BglII.
Para cada linea de clavel diferentes cantidades de ADN precalentado (10, 7, 5 y 2 pg) se digirieron durante 12 h con una de estas enzimas de restriccion BamHI, BglII, EcoRI o HindIII.
2002 and by restriction analysis using the BglII restriction enzyme as recommended by the manufacturer (Amersham Biosciences, Athens, Greece).
We used BglII, as outlined in the methods below, which cuts once inside the vector and also once in LC3.
For this ribozyme-specific primers were designed and synthesized to include HindIII and BglII restriction sites: Vif5113BglII Forward (5' ATT AGA TCT ACA TAT GGT GTT TCT GAT GAG 3') and Vif5113HindIII Reverse (5' ATT AAG CTT ATG GAA AAG ATT AGT TTC G 3').