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인용 및 참조 문헌 (18)
Invitrogen™
Zero Blunt™ PCR Cloning Kit
The Zero Blunt™ PCR Cloning Kit offers an easy method for high-efficiency (>80%) cloning of blunt-end PCR products amplified with자세히 알아보기
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보기 방식 변경
| 카탈로그 번호 | 수량 |
|---|---|
| K270020 | 20 Reactions |
| K270040 | 40 Reactions |
카탈로그 번호 K270020
제품 가격(KRW)
757,000
Online offer
Ends: 31-Dec-2025
890,000할인액 133,000 (15%)
Each
수량:
20 Reactions
제품 가격(KRW)
757,000
Online offer
Ends: 31-Dec-2025
890,000할인액 133,000 (15%)
Each
The Zero Blunt™ PCR Cloning Kit offers an easy method for high-efficiency (>80%) cloning of blunt-end PCR products amplified with proof-reading, thermostable DNA polymerases. The Zero Blunt™ PCR Cloning Kit uses the multipurpose cloning vector pCR™-Blunt and ExpressLink™ T4 DNA Ligase to generate a ligation product in a five-minute, room-temperature ligation step.
Features of the Zero Blunt™ Cloning™ Kit with pCR™-Blunt vector:
• Fast & convenient—5-minute, room-temperature ligation
• Efficient—ccdB gene for positive selection results in >80% clones with correct insert
• Flexible—choice of kanamycin or Zeocin™ resistance for flexible antibiotic selection
The pCR™-Blunt vector provides:
• EcoR I sites flanking the PCR product insertion site for excision of inserts
• T7 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening
How Zero Blunt™ PCR Cloning Works
The Zero Blunt™ PCR Cloning Kit is designed to clone blunt PCR fragments (or any blunt DNA fragment) with a low background of non-recombinants. The pCR™-Blunt vector contains the lethal E. coliccdB gene fused to the C-terminus of LacZα (Bernard et al., 1994). Ligation of a blunt PCR fragment disrupts expression of the lacZα-ccdB gene fusion permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed when the transformation mixture is plated.
Kit Configurations
The Zero Blunt™ PCR Cloning Kit is offered in a variety of configurations: with One Shot™ TOPO10 Chemically Competent E. coli (K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40) in 20- and 40- reaction kit sizes.
Features of the Zero Blunt™ Cloning™ Kit with pCR™-Blunt vector:
• Fast & convenient—5-minute, room-temperature ligation
• Efficient—ccdB gene for positive selection results in >80% clones with correct insert
• Flexible—choice of kanamycin or Zeocin™ resistance for flexible antibiotic selection
The pCR™-Blunt vector provides:
• EcoR I sites flanking the PCR product insertion site for excision of inserts
• T7 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening
How Zero Blunt™ PCR Cloning Works
The Zero Blunt™ PCR Cloning Kit is designed to clone blunt PCR fragments (or any blunt DNA fragment) with a low background of non-recombinants. The pCR™-Blunt vector contains the lethal E. coliccdB gene fused to the C-terminus of LacZα (Bernard et al., 1994). Ligation of a blunt PCR fragment disrupts expression of the lacZα-ccdB gene fusion permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed when the transformation mixture is plated.
Kit Configurations
The Zero Blunt™ PCR Cloning Kit is offered in a variety of configurations: with One Shot™ TOPO10 Chemically Competent E. coli (K2700-20 and K2700-40) and without competent cells (K2750-20 and K2750-40) in 20- and 40- reaction kit sizes.
For Research Use Only. Not for use in diagnostic procedures.
사양
박테리아 또는 효모 균주TOP10
클로닝 방법Blunt PCR
제품라인Zero Blunt
제품 유형PCR Cloning Kit
프로모터T7
수량20 Reactions
벡터Blunt DNA Cloning Vectors
형식Kit
Unit SizeEach
구성 및 보관
Zero Blunt™ PCR cloning kits contain linearized pCR™-Blunt vector, ExpressLink™ T4 DNA ligase , 5X ExpressLink™ T4 DNA ligase buffer, control template, dNTPs, sterile water, and M13 forward and reverse primers. Competent cell kits contain One Shot™ chemically competent E. coli, S.O.C. medium, and a supercoiled control plasmid.
Store the One Shot™ E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.
Store the One Shot™ E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.
자주 묻는 질문(FAQ)
What is the best molar ratio of PCR product:vector to use for TOPO TA cloning? Is there an equation to calculate the quantity to use?
What is the best ratio of insert:vector to use for cloning? Is there an equation to calculate this?
I'm seeing a lot of vector-only colonies when I try to perform a negative control reaction using vector only (no insert) for a TOPO reaction. Is my TOPO vector re-ligating?
I'm trying to clone in my phosphorylated PCR product into a TOPO vector, and I'm getting no colonies. However, when I clone the same product into a TA vector, everything works perfectly. Why is this?
I'm able to get a lot of colonies, however, none contain my insert of interest. What should I do?
인용 및 참조 문헌 (18)
인용 및 참조 문헌
Abstract
Utilization of MHC class I transgenic mice for development of minigene DNA vaccinesencoding multiple HLA-restricted CTL epitopes.
Journal:J Immunol
PubMed ID:10201910
'We engineered a multiepitope DNA minigene encoding nine dominant HLA-A2.1- and A11-restricted epitopes from the polymerase, envelope, and core proteins of hepatitis B virus and HIV, together with the PADRE (pan-DR epitope) universal Th cell epitope and an endoplasmic reticulum-translocating signal sequence. Immunization of HLA transgenic mice with this construct
Interactions between protein kinase CK2 and Pin1. Evidence for phosphorylation-dependent interactions.
Journal:J Biol Chem
PubMed ID:11940573
'The peptidyl-prolyl isomerase Pin1 interacts in a phosphorylation-dependent manner with several proteins involved in cell cycle events. In this study, we demonstrate that Pin1 interacts with protein kinase CK2, an enzyme that generally exists in tetrameric complexes composed of two catalytic CK2 alpha and/or CK2 alpha'' subunits together with two
Role of CCAA nucleotide repeats in regulation of hemoglobin and hemoglobin-haptoglobin binding protein genes of haemophilus influenzae.
Journal:J Bacteriol
PubMed ID:10482534
'Haemophilus influenzae utilizes hemoglobin and hemoglobin-haptoglobin as heme sources. The H. influenzae hemoglobin- and hemoglobin-haptoglobin binding protein genes, hgpA, hgpB, and hgpC, contain lengths of tetrameric CCAA repeats. Using an hgpA-lacZ translational gene fusion, we demonstrate phase-variable expression of lacZ associated with alteration in the length of the CCAA repeat
Regulation of the Bub2/Bfa1 GAP complex by Cdc5 and cell cycle checkpoints.
Journal:Cell
PubMed ID:11733064
'During mitosis, a ras-related GTPase (Tem1) binds GTP and activates a signal transduction pathway to allow mitotic exit. During most of the cell cycle, Tem1 function is antagonized by a GTPase-activating protein complex, Bfa1/Bub2. How the Bfa1/Bub2 complex is regulated is not well understood. We find that Polo/Cdc5 kinase acts
Substantially enhanced cloning efficiency of SAGE (Serial Analysis of Gene Expression) byadding a heating step to the original protocol.
Journal:Nucleic Acids Res
PubMed ID:9889294
'The efficiency of the original SAGE (Serial Analysis of Gene Expression) protocol was limited by a small average size of cloned concatemers. We describe a modification of the technique that overcomes this problem. Ligation of ditags yields concatemers of various sizes. Small concatemers may aggregate and migrate with large ones