transbionovo

+86-10-57815027      
中文版 | ENGLISH

Your present location:HomeProductsCloning and Mutagenesis SystemCloning Vectors / pEASY®-T1 Simple Cloning Kit
  • Product Name: pEASY®-T1 Simple Cloning Kit
  • Catalog Number: CT111
  • Storage: Competent Cell at -70°C for six months; others at -20°C for one year
  • 币种:  
    • Specification: 20 rxns 60 rxns
  • Quantity:  

Quick purchase Purchase Favorite

 

Overview Contents& storage Citations & references Related Images Download  
Overview

Description

pEASY ®- T1 Simple Cloning Vector eliminates the multi-cloning sites of pEASY ®-T1 Cloning Vector. It is designed for cloning and sequencing Taq-amplified PCR products.

•5 minutes fast ligation of Taq-amplified PCR products.

•Kanamycin and Ampicillin resistance genes for selection.

•Easy blue/white selection.

•SR primer and M13 forward primer for sequencing.

•T7 promoter for in vitro transcription.

•Trans1-T1 Phage Pesistant Chemically Competent Cells, high transformation efficiency (>109 cfu/μg pUC19 DNA) and fast growing.

 

Preparation

1.Primer requirement: primer cannot be phosphorylated

2.PCR Enzyme: Taq DNA polymerases

3.Reaction conditions: in order to ensure the integrity of amplification products, 5-10 minutes of post-extension step is required. After amplification reaction, use agarose gel electrophoresis to verify the quality and quantity of PCR product

 

Reaction System

Add following components into a microcentrifuge tube.

PCR products 0.5-4 μl (can be increased or reduced based on PCR product yield, no more than 4 μl)

pEASY ®- T1 Cloning Vector 1 μl

Gently mix well, incubate at room temperature (20oC-37oC) for 5 minutes. After reaction, place the tube on ice.

1. Optimal amount of insert

Molar ratio of vector to insert = 1:7 (1 kb, ~20 ng; 2 kb, ~40 ng)

2.Optimal volume of vector: 1 μl(10 ng)

3.Optimal reaction volume: 3~5 μl

4.Optimal incubation time

(1)0.1~1 kb (including 1 kb): 5~10 minutes

(2)1~2 kb (including 2 kb): 10~15 minutes

(3)2~3 kb (including 3 kb): 15~20 minutes

(4)≥3 kb: 20~30 minutes

Use the maximum incubation time if the insert is gel purified.

5.Optimal incubation temperature: for most PCR inserts, the optimal temperature is about 25oC; for some PCR inserts, optimal results can be achieved with higher temperature (up to 37oC).

 

Transformation

1.Add the ligated products to 50 μl of Trans1-T1 Phage Resistant Chemically Competent Cell and mix gently (do not mix by pipetting up and down).

2.Incubate on ice for 20~30 minutes.

3.Heat-shock the cells at 42oC for 30 seconds.

4.Immediately place the tube on ice for 2 minutes.

5.Add 250 μl of room temperature SOC or LB medium. Shake the tube at 37oC (200 rpm) for 1 hour.

6.In the meantime, mix 8 μl of 500 mM IPTG with 40 μl of 20 mg/ml X-gal. Spread them evenly onto a selective LB plate. Place the plate at 37oC for 30 minutes.

7.Spread 200 μl or all transformants on the pre-warmed plate. Incubate at 37oC overnight.

 

Contents& storage

 

Component

CT111-01 (20 rxns)

CT111-02 (60 rxns)

pEASY ®-T1 Simple Cloning Vector (10 ng/μl)

20 µl

3×20 µl

Control Template (5 ng/μl)

5 µl

5 µl

Control Primers (10 μM)

5 µl

5 µl

M13 Forward Primer (10 μM)

50 µl

150 µl

M13 Reverse Primer (10 μM)

50 µl

150 µl

SR Primer (10 μM)

50 µl

150 µl

Trans1-T1 Phage Resistant Chemically Competent Cell

10×100 μl

30×100 μl

 

Citations & references
Literature Journal IF Author Date Link
T-2 toxin is hydroxylated by chicken CYP3A37 Food and Chemical Toxicology 3.01 Yuan Y, et al. 2013 Sep http://www.ncbi.nlm.nih.gov/pubmed/24090734
A comparison of microbial communities in deep-sea polymetallic nodules and the surrounding sediments in the Pacific Ocean Deep Sea Research Part I: Oceanographic Research Papers 2.816 Wu Y H, et al. 2013 Sep http://www.sciencedirect.com/science/article/pii/S0967063713000940
Characterization of chicken cytochrome P450 1A4 and 1A5: Inter-paralog comparisons of substrate preference and inhibitor selectivity Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2.707 Yang J, et al. 2013 May http://www.ncbi.nlm.nih.gov/pubmed/23474502
Plasmid-mediated quinolone resistance genes and antibiotic residues in wastewater and soil adjacent to Swine feedlots: potential transfer to agricultural lands Environmental Health Perspectives 7.26 Li J, et al. 2012 Aug http://www.ncbi.nlm.nih.gov/pubmed/22569244
Molecular Characterization and Expression Pattern of Tripartite Motif Protein 39 in Gallus gallus with a Complete PRY/SPRY Domain International Journal of Molecular Sciences 2.598 Pan C, et al. 2011 http://www.ncbi.nlm.nih.gov/pubmed/21747707
Trends in antibiotic resistance genes occurrence in the Haihe River, China Environmental Science and Technology 5.257 Luo Y, et al. 2010 Oct http://www.ncbi.nlm.nih.gov/pubmed/20509603
Nitrogen deposition mediates the effects and importance of chance in changing biodiversity Molecular Ecology 6.275 Ximei Zhang, et al. 2011 Jan

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-294X.2010.04933.x/abstract;jsessionid=95DB091914EC73F67071

CB233DEFB98F.f02t02?deniedAccessCustomisedMessage

=&userIsAuthenticated=false

Allelic Variants at the TaZds-D1 Locus on Wheat Chromosome 2DL and their Association with Yellow Pigment Content ACSESS   Caiying Zhang, et al. 2011 Jul https://dl.sciencesocieties.org/publications/cs/abstracts/51/4/1580
Antibacterial and antiviral properties of tongue sole (Cynoglossus semilaevis) high mobility group B2 protein are largely independent on the acidic C-terminal domain Fish & Shellfish Immunology 2.964 Hao Long, et al. 2014 Mar http://www.sciencedirect.com/science/article/pii/S105046481400014X
Rock bream (Oplegnathus fasciatus) IL-12p40: Identification, expression, and effect on bacterial infection Fish & Shellfish Immunology 2.964 Lu Zhang, et al. 2014 Aug http://www.sciencedirect.com/science/article/pii/S1050464814001752
Cloning of seed dormancy genes (TaSdr) associated with tolerance to pre-harvest sprouting in common wheat and development of a functional marker Theoretical and Applied Genetics 3.658 Yingjun Zhang, et al. 2014 April http://link.springer.com/article/10.1007/s00122-014-2262-6
TaGS-D1, an ortholog of rice OsGS3, is associated with grain weight and grain length in common wheat Molecular Breeding 3.251 Yingjun Zhang, et al. 2014 May http://link.springer.com/article/10.1007/s11032-014-0102-7
Identification and biochemical characterization of five long-chain acyl-coenzyme A synthetases from the diatom Phaeodactylum tricornutum Plant Physiology and Biochemistry 2.775 Xiaojing Guo, et al. 2014 Jan http://www.sciencedirect.com/science/article/pii/S0981942813003860
Isolation and characterization of twelve novel polymorphic tetranucleotide microsatellite loci for Tsinling Tree Toad (Hyla tsinlingensis) Conservation Genetics Resources 1.172 Xing Kang, et al. 2015 Mar http://link.springer.com/article/10.1007/s12686-015-0445-3
CD83 is required for the induction of protective immunity by a DNA vaccine in a teleost model Developmental & Comparative Immunology 2.815 Mo-fei Li, et al. 2015 Mar http://www.sciencedirect.com/science/article/pii/S0145305X15000580
Development and application of a novel screening method and experimental use of the mutant bacterial strain Clostridium beijerinckii NCIMB 8052 for production of butanol via fermentation of fresh cassava RSC Advances 3.84 Haifeng Su, et al. 2015 Jan http://pubs.rsc.org/en/content/articlelanding/2015/ra/c4ra16576d#!divAbstract
CsCTL1, a teleost C-type lectin that promotes antibacterial and antiviral immune defense in a manner that depends on the conserved EPN motif Developmental & Comparative Immunology 2.815 Ze-jun Zhou, et al. 2015 Jan http://www.sciencedirect.com/science/article/pii/S0145305X15000142
Akt‑mediated phosphorylation of Oct4 is associated with the proliferation of stem‑like cancer cells Oncology
Reports
2.301 Qing-Wei Zhao, et al. 2015 Jan http://www.spandidos-publications.com/or/33/4/1621?text=fulltext
Reconstructed metagenomes reveal changes of microbial functional profiling during PAHs degradation along a rice (Oryza sativa) rhizosphere gradient Journal of Applied Microbiology 2.479 B. Ma, et al. 2015 Jan

http://onlinelibrary.wiley.com/doi/10.1111/jam.12756/abstract?systemMessage

=Wiley+Online+Library+will+be+disrupted+on+11th+July+2015+at+10%3A00

-16%3A00+BST+%2F+05%3A00-11%3A00+EDT+%2F+17%3A00-23%3A00++SGT++for+essential+maintenance.++Apologies+for+the+

inconvenience

Long-term application of organic manure changes abundance and composition of ammonia-oxidizing archaea in an acidic red soil Soil Science and Plant Nutrition 0.729 Weishou Shen, et al. 2015 Mar http://www.tandfonline.com/doi/abs/10.1080/00380768.2015.1023687#.VaWxSPmqqko
P247 and P523: Two In Vivo-Expressed Megalocytivirus Proteins That Induce Protective Immunity and Are Essential to Viral Infection PLoS One 3.73 Jian Zhang, et al. 2015 Jan http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376877/
RcRR1, a Rosa canina Type-A Response Regulator Gene, Is Involved in Cytokinin-Modulated Rhizoid Organogenesis PLOS ONE 3.73 Bin Gao,et al. 2013 Aug http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0072914
Related Images

  To be closed automatically in X seconds

View my favorites

Related products
no such related products!