We found that BLM greatly enhances the random integration of transfected plasmids into human cells. Importantly, this enhancement was independent of the molecular form of the plasmid, the cell type or the transfection method, suggesting that the BLM effect is intrinsically general. Transient expression...
The donor plasmid contains Tet2 guide cleavage sites on either side of the transgene but does not have any sequences that are homologous to the AAVS integration site. B Overview of the knock-in assay. Three plasmids are transfected into HEK293 cells (i) Cas9 (wt...
Chromosomal DSB joining assay. An all-in-one CRISPR vector, pX330-U6- Chimeric_BB-CBh-hSpCas9 (pX330, Addgene plasmid #42230), was used to express Cas9 and single-guide RNA. Sense and antisense oligonucleotides of each single-guide RNA targeting sequence were annealed and cloned into pX...
Random mutagenesis, such as error-prone PCR (epPCR), is a technique capable of generating a wide variety of a single gene. However, epPCR can produce a large number of mutated gene variants, posing a challenge in ligating these mutated PCR products into plasmid vectors. Typically, the primer...
This may indicate the loss of the plasmid in absence of selection pressure. The intracellular survival experiment has also conclusively indicated a reversal of the mutation. The complemented strain showed more similar growth tendency towards wild-type strain than towards the mutant (Figure 7 B). ...
A quantitative assay for HIV DNA integration in vivo 热度: sperm cells and foreign dna a controversial relation 热度: RESEARCHARTICLEOpenAccess Homologousillegitimaterandomintegrationof foreignDNAintotheXchromosomeofa transgenicmouseline BowenYan 1
Excitable systems of eukaryotic chemotaxis can generate asymmetric signals of Ras-GTP-enriched domains spontaneously to drive random cell migration without guidance cues. However, the molecules responsible for the spontaneous signal generation remain elu
(15 s at 94 °C + 30 s at 60 °C + 30 s at 72 °C) × 40. Absolute quantification of nuclease-resistant viral genomes was performed using serial dilutions of standards with known copy number as measured by Quant-iT™ PicoGreen® dsDNA Assay and NanoDrop™1000 (...
QIAGEN 1054575 HiSpeed Plasmid Giga EF Kit (5) QIAGEN 1061072 AIV-H5 RT-PCR REAGENT QIAGEN 1061094 HCV RT-PCR REAGENT VII QIAGEN 1061115 PG FMDV RT-PCR Assay (48) Z QIAGEN 9016711 Security Key, LC Analyzer, encoded QIAGEN 9016685 Washer Mechanical, RCS1 ...
pMIN_VP3 was created by excising VP3 from pRepCap_RR using Nco I and BfuA I sites, and ligating to a minimized plasmid backbone generated by sequence overlap extension48 using pBAD24 as a template. pInSALect_RR was created by removal of RaPID restriction sites from pInSALect, followed ...