CRISPR cas9 RNP

HT115(DE3) Competent cells are usable for RNAi interference assays.

Cat: T-C3760

Size: 20×100μL

Price: 197,60 €

Storage: Store at -70°C, valid for 6 months.（Avoid repeated freezing and thawing）

Genotype: F-mcrA mcrB, IN(rrnD-rrnE)1 rnc14::Tn10λ(DE3 lavUV5::T7 polymerase)

Strain Resistance: Sensitive to ampicillin, kanamycin, chloramphenicol, and

streptomycin. Has tetracycline resistance.

Introduction:

HT115(DE3) is an RNase III defective Escherichia coli strain that can be expressed by IPTG

induction to produce dsRNA. It is used for RNAi interference assay. The strain also has the DE3

region of λ phage, which can express T7 RNA polymerase, and is suitable for the efficient

expression of prokaryotic expression vectors containing T7 promoters (such as pET, etc.). Non-T7

promoter expression vectors (such as pGEX, pMal, pTrc, etc.) can also be expressed in this strain.

HT115(DE3) competent cells were prepared by a special process. The conversion efficiency of

pUC19 plasmid was 1×106 cfu/μg DNA.

T7E1 enzyme cleaves heteroduplex DNA only.

Using the genomic DNA extracted from cells or tissues subjected to gene-editing processes as template, target DNA region that possibly contains mutations is amplified by High-Fidelity DNA polymerase. The PCR products are then denatured and annealed, followed by digestion with T7E1 enzyme which cleaves heteroduplex DNA only.

Genome-Editing Mutation Detection Kit provides a simple, reliable, and rapid method for detecting the mutations on genomic DNA extracted from cells or tissues subjected to CRISPR/Cas9 or other gene-editing processes. It employs the High-Fidelity DNA polymerase for amplifying the target DNA fragments, and the T7 Endonuclease I (T7E1) for recognizing and cleaving non-perfectly matched DNA (heteroduplex DNA).

Genome-Editing Mutation Detection Kit contains all reagents for DNA extraction, PCR amplification with high fidelity, and T7E1 enzyme digestion. It also contains positive control template DNA and its corresponding primers for troubleshooting. The DNA extracts and PCR products can be added directly into the subsequent reactions without extra purification, enabling the kit convenient to use.

Working mechanism of Genome-Editing Mutation Detection Kit: Using the genomic DNA extracted from cells or tissues subjected to gene-editing processes as template, target DNA region that possibly contains mutations is amplified by High-Fidelity DNA polymerase. The PCR products are then denatured and annealed, followed by digestion with T7E1 enzyme which cleaves heteroduplex DNA only. If mutations exist in target DNA, heteroduplex will be formed between unmutated DNA and mutated DNA, or between mutated DNA with different mutations. The mutated DNA is then cleaved by T7E1 into smaller fragments. Based on the concentrations of cleaved and uncleaved fragments, the efficiency of genome editing can be estimated.

Control Template provided in this kit contains both unmutated and mutated gene fragments. Amplicons of unmutated fragments are 525bp in length, and those of mutated fragments with 7nt deletion mutation are 518bp in length. The mutated DNA strand in the Heteroduplex DNA formed between the unmutated and the mutated DNA fragments is cleaved by T7E1 into two DNA fragments, 288bp and 230bp in length, respectively. Meanwhile, the homoduplex of either unmutated or mutated DNA could not be cleaved by T7E1. Therefore, after T7E1 digestion, positive control samples should contain DNA fragments of four different sizes (525bp, 518bp, 230bp, and 288bp). Usually, the 525bp and 518bp bands are nearly together when separated on agarose gels.


Katalog Nummer: 25-6050 DNA

Gene Symbol : USP47 // Species: human

NCBI Gene ID : 55031

Ensembl ID: ENSG00000170242 // LINK : https://www.ensembl.org/index.html?redirect=no

Uniprot ID: Q96K76

Chemically synthesized sgRNA (300pmol), LM RNP-specific transfection reagent, LM cell lysate, PCR primers, and sequencing primers.

10 reactions /1Target gene，

1Target gene /1.5mL tube

High Efficiency Knockout – Utilizes chemically synthesized and modified sgRNAs with Cas9 RNPs for rapid and precise USP47 gene disruption.

Targeted Early Exon Cleavage – Ensures loss-of-function by fragment deletion in early exons.

Ready-to-Use Kit – Includes all necessary components: sgRNA, RNP transfection reagent, cell lysate, PCR and sequencing primers.

Minimal Off-Target Effects – Optimized sgRNA design reduces unintended gene editing.

Flexible Storage – Long-term storage at -80℃ (1 year) or short-term at -20℃ (6 months) with minimal activity loss.

Human eukaryotic cell gene knockout experiments in vitro

Functional genomics research

Disease mechanism studies

Drug target validation

Cell line engineering for research

Specification Verification: All sample parameters are cross-checked, including biological characteristics, container type, and documentation.

Quality Assurance: Manufactured under ISO 13485 standards with complete batch traceability for reproducible research.

Reliable & Traceable: Validated for consistent performance across experiments, ensuring safe and dependable use.

We offer bulk supply and wholesale USP47 CRISPR Easy KO Kit options for research institutes, laboratories, and distributors.

Benefits include:

Competitive pricing for large-volume orders

Consistent batch quality for multiple experiments

Fast shipping with temperature-controlled logistics

Dedicated support for training and technical queries

Get in touch with us now to talk about your inquiries.

Please talk with Klaus

+49 (0)1575 807 6839

klaus@sansunbiologics.com

FOR RESEARCH USE ONLY

"ready-to-use" : USP30 ** / USP47 / TPP2 / HLA-DQB1 uvm. KO kits available.
please inquire : info@sansunbiologics.com