증폭 및 분석장비	추출 및 분주장비	기초 및 일반장비
유전자 분석장비 Nanostring nCounter System POCKIT Smart PCR System UV-VIS Spectrophotometer/Fluorometer Mic real 이미지 분석장비 Enduro Gel-Doc Enduro GDS Touch Azure Imaging System Sapphire Biomolecular Imager 계수기 Moxi Z Mini Automated Cell Counter Moxi Go II CellDrop™ Cell Counters Moxi GO 532 유전자 증폭장비 SuperCycler Microplate reader Ao Absorbance Microplate Reader Byonoy Absorbance 96 LSPR 분석기 Instant NanoBiosensors INB-D200 Biomarker Analyzer	추출장비 taco Aurora System Semi-Automated Nucleic Acid Extraction System taco Mini Automatic Nucleic Acid Extraction System iColumn Extraction System iColumn LV System 자동화 장비 BlotCycler™ 조직파쇄기 tacoPrep Bead beater	소형원심분리기 Multi-Well Plate Centrifuge 12 strip tube Centrifuge SmartFuge Centrifuge 전기영동장치 Protein Electrophoresis System Blotting System vortex MTV-1 mini vortex Lab instrument LED Transilluminator Rotary Mixing Solution CO-01 Tube Roller CO-02/CO-02S Vacuum Pump CO-03 / CO-03S Mini Stirring Magnetic (Cute Stir) CO-12 Digital Magnetic Stirrer Hot Plate Series CO-04/CO-05/CO-05S Glass Beads Sterilizer CO-06 Handheld Homogenizer CO-07 Handheld UV Analyzer CO-08 Mini Handheld Homogenizer CO-11 Black-box Type UV Analyzer CO-09 Mupid Mini-EP Mini Electrophoresis System

Advansta	Epicentre	기타제품
Fluorescent Western Blotting MultiColor Fluorescent Secondary Antibodies Streptavidin Conjugates FLASHBlot Transfer Buffer Chemiluminescent Western Blotting WesternBright ECL & ECL Spray HRP Conjugates Quantum HRP Substrate Sirius HRP Substrate WesternBright ChemiPen ELISA HRP Conjugates ELISABright Electrophoresis Protein sample loading buffers Scarlet protein stain Purification SDS-PAGE sample preparation kit RapidClean Buffers and Solutions blocking solution Washing solution Avant buffer pouches LucentBlue X-ray film Background quenching sheets Transfer membranes Incubation trays Blotting papers Visio	Array-Based Analysis RNA Amplification for qPCR RNA Labeling RNA Labeling for Expression Microarrarys End-Point PCR & RT End-Point PCR Kits cDNA Synthesis dNTP Solutions Nucleic Acid Purification DNA Extraction RNA Extraction DNA Purification. Genomic DNA Purification. Cloned RNA Purification Lysozymes In Vitro Transcription In Vitro Transcription Kits NTP Solutions Library Construction Vector and Insert Preparation Cloning, cDNA Cloning, Genetics Competent Cells Additional Reagents Transposomics™ In Vivo Transposomics™ In Vitro Transposomics™ RNA Polyadenylation RNA Polyadenylation	Instant-Bands Phytotech Phyto Tech 제품 Ginrei Lab Interactive Co-Culture Plate (ICCP) Synthego Engineered Cells

Applications	Product Catalog	NEB 제품 견적 및 주문	NEBNext - NGS Kit
Cellular Analysis Cloning & Synthetic Biology DNA Amplification, PCR & qPCR Genome Editing RNA Analysis Sample Preparation for Next Generation Sequencing Epigenetics Protein Expression & Purification Protein Analysis & Tools Glycobiology & Proteomics	Restriction Endonucleases PCR, qPCR & Amplification Technologies DNA Modifying Enzymes Sample Preparation for Next Generation Sequencing Nucleic Acid Purification Markers & Ladders RNA Reagents DNA Assembly, Cloning and Mutagenesis Kits Genome Editing Gene Expression Cellular Analysis Epigenetics Protein Expression & Purification Technologies Competent Cells Protein Tools Glycobiology DNA Plasmids & Substrates Buffers Strains Discontinued	NEB 제품 견적 및 주문 문의 NEB 샘플 신청 NEB Info.	NEBNext Direct® Cancer HotSpot Panel NEBNext® rRNA Depletion Kit NEBNext® Ultra™ II RNA Library Prep Kit NEBNext® Microbiome DNA Enrichment Kit NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® NEBNext Direct® BRCA1BRCA2 Panel NEBNext® small RNA Library Prep Kit

Purification	Real Time PCR Enzyme	PCR & RT Enzyme	Lab Essential
Genomic DNA eCube Tissue DNA Mini Kit eCube Plant DNA Mini Kit eCube Stool DNA Mini Kit eCube Soil DNA Mini Kit eCube Blood DNA Mini Kit RNA eCube Blood RNA Mini Kit eCube Tissue RNA Mini Kit eCube Plant RNA Mini Kit eCube RNA Clean-up Kit eCube-Zol RNA Reagent eCube-RNA Stabilization Solutions Virus NA eCube Viral Nucleic Acid Mini Kit Plasmid eCube Plasmid DNA Mini Kit eCube Plasmid DNA Midi/Maxi Kit Gel & PCR purification eCube PCR Purification Kit eCube Gel&PCR Purification Kit eCube Gel Purification Kit	1-Step QuantiSpeed Probe One-Step Kit QuantiSpeed SYBR One-Step Kit 2-Step QuantiSpeed Probe Kit QuantiSpeed SYBR Green Kit HRM QuantiSpeed HRM Kit QuantiSpeed Genotyping Kit	제품 준비중	RNase Inhibitor Agarose Agarose dNTP dNTP Solutions DNA ladder & Loading dye DNA Loading Dye DNA Ladder

nCounter 서비스	Real Time PCR 서비스
nCounter 서비스	Real Time PCR 서비스

CopyCutter^TM, CopyControl^TM

Gene and PCR Cloning Kit

CopyCutter™ EPI400™ Electrocompetent E. coli
CopyCutter™ EPI400™ Chemically Competent E. coli

Applications

Cloning of unstable DNA sequences or those expressing toxic proteins.

CopyCutter™ EPI400™ E. coli* cells were developed to significantly lower the copy number of a wide variety of common vectors so that you can more readily clone unstable DNA sequences. DNA that is unstable at high-copy number often codes for a protein that inhibits cell growth or contains AT- and GC-rich sequences or sequences with strong secondary structure (Fig. 2).¹

The CopyCutter EPI400 cell line was derived from Epicentre's high-transformation efficiency phage T1-resistant TransforMAX™ EC100™-T1^R E.coli strain by manipulating a gene that controls the copy number of vectors containing ColE1 or pMB1 origins of replication (e.g., pUC- and pET-type vectors). This constitutively expressed gene, pcnB (plasmid copy number), was deleted from the TransforMAX EC100 strain and replaced with a modified pcnB gene that is linked to an inducible promoter, creating the CopyCutter EPI400 strain.

The copy number of ColE1-type vectors in the CopyCutter EPI400 strain compared to the parental TransforMAX EC100 strain is approximately 4- to 25-fold lower, depending on the vector. Moreover, a short incubation in the presence of the CopyCutter Induction Solution can increase the copy number of the vector to improve plasmid yields (Fig. 3).

Figure 1. Copy-number of ColE1-type plasmids is lowered up to 25-fold in CopyCutter™ EPI400™ E. coli cells. Lanes C, TransforMax™ EC100™ cells; Lanes U and I, uninduced or induced CopyCutter EPI400 cells. DNA extracts from the same number of lysed cells (based on OD₆₀₀) were loaded per lane.

Figure 2. DNA inserts encoding toxic gene products were successfully cloned into high-copy-number vectors using CopyCutter™ EPI400™ E. coli cells. After sequencing, the full-length acpP clones in TransforMAX™ EC100™ cells were found to contain multiple point mutations.

Figure 3. Uninduced CopyCutter™ EPI400™ E. colicells containing a regB clone (lane U) are induced to higher-copy number (lane I) using the CopyCutter Induction Solution. Crude extracts of plasmid DNA were prepared from cells grown in selective media and analyzed by agarose gel electrophoresis. Approximately the same number of lysed cells (based on A₆₀₀) were loaded per lane.

Benefits

Maintain clones at low-copy number, then induce to higher copy number for improved plasmid yield.

High transformation efficiency with clones of all sizes.

tonAfor resistance to bacteriophages T1 and T5.

lacZΔM15for blue/white screening of recombinants.

Restriction minus [mcrA, Δ(mrr-hsdRMS-mcrBC)] genotype enables efficient cloning of methylated DNA.

Endonuclease minus (endA1) to ensure high yields of DNA.

Recombination minus (recA1) for greater stability of large cloned inserts.

Genotype
F^- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ^- rpsL (Str^R) nupG trfA tonA pcnB4 dhfr

CopyCutter EPI400 Electrocompetent E. coli

Transformation efficiency of >1 x 10¹⁰ cfu/µg of pUC19.

CopyCutter EPI400 Chemically Competent E. coli

Transformation efficiency of >1 x 10⁷ cfu/µg of pUC19.

Reference

Haskins, D. (2004) Epicentre Forum 11(5), 6.

*Covered by issued and/or pending patents.

CopyControl™ cDNA, Gene and PCR Cloning Kits

Applications

Efficient cloning of any insert up to 15 kb, including those that may be unstable or encode proteins that are toxic to the E. coli host cells.

Up to ~25% more cDNA clones can be obtained in a library if the cDNA clones are present in the host cell at only about one copy per cell (P. Carninci and W. Szybalski, personal communication), presumably due to reduced detrimental effects on host cell metabolism and lower toxicity of expressed clone sequences and gene products. The CopyControl™ cDNA, Gene, and PCR Cloning Kits* provide a rapid and improved method (Fig. 1) for cloning any blunt-end DNA fragments (e.g., genomic DNA, PCR products, or cDNA) at single-copy number, with the capability to induce to high-copy numbers as needed (Fig. 2). The CopyControl pCC1™ (Blunt) Vector provided in the kits contains both the E. coli F-factor single-copy origin of replication (oriII) and an inducible high-copy oriV. Replication does not occur using the oriV unless and until the trfA gene in the chromosome of TransforMax™ EPI300™ cells (which is under the control of a tightly regulated promoter) is induced. Thus, CopyControl cDNA, PCR, or gene clones can be grown at single copy to ensure insert stability and successful cloning of sequences that are toxic to the host cell. Then, any desired clone can be induced up to 200 copies per cell for high yields of DNA for all downstream applications.

Benefits

Obtain more cDNA or PCR clones and more complete clone libraries (Table 1).

Obtain fewer mutated clones. Clones having mutations in genes that are toxic to the host cell are selectively favored at high-copy number.

Screen clones for insert size in 1 hour or less without the need for overnight cultures or restriction endonuclease digests (Fig. 3).

Figure 1 . Efficient, blunt-end cloning of inserts up to 15 kb and screening of clones can be completed in 24 hours using the CopyControl™ cDNA, Gene, and PCR Cloning Kit. Clones can be kept at one copy per cell for clone stability and then, whenever desired, any clone can be induced to high-copy number (up to 200 copies per cell) for high yields of DNA.

Table 1. PCR product cloning results with the CopyControl™ System.

Figure 2. Single-copy CopyControl™ clones can be induced to 200 copies per cell. DNA from an equal number of uninduced (–) cells (lane 1) and undiluted induced (+) cells (lane 5) containing a 10-kb CopyControl PCR clone was analyzed by electrophoresis in a 1% agarose gel. Copy number in induced cells was estimated by visually comparing the staining intensity of the diluted induced sample with the uninduced sample. DNA from induced cells was diluted 1:200 (lane 2), 1:100 (lane 3), and 1:50 (lane 4).

Figure 3. Screening of clones from a 5-kb PCR product. The PCR product was cloned using the CopyControl™ cDNA, Gene, and PCR Cloning Kit. Very small portions from nine white colonies were screened using the kit, confirming the presence of the 5-kb insert. Lane M, DNA size ladder; lane 1, 8-kb supercoiled DNA marker; lanes 2-10, randomly chosen clones. Total time for colony screening, including gel electrophoresis, was approximately 60 minutes.

*Covered by issued and/or pending patents.

CopyCutterTM, CopyControlTM

Gene and PCR Cloning Kit

CopyCutter^TM, CopyControl^TM