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TOPO™ TA Cloning™ Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction, and are available with a variety of competent cells, or no competent cells, depending on your needs and budget. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice. Features of TOPO® TA Cloning® Kits for Sequencing:

Get more sequence—allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations

TOPO® TA Cloning® Kits for Sequencing—overview

Vector: pCR™4-TOPO® TA vector—optimized cloning vector for improved sequencing results

Cloning method: TOPO® TA Cloning®—Topoisomerase I–based, 5-minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector

Competent cells: Various options—choose from kits with general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells, or use your own

pCR™4-TOPO® TA vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA clone selection and manipulation
The pCR™4-TOPO® TA vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection and blue/white screening. The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.

Simplified TOPO®-based cloning
Using TOPO® cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform with the providedE. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones and save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products, which contain 3´A overhangs.

The standard in cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

TOPO® TA Cloning® Kits for Sequencing—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)
• Provide your own: for flexibility and to save money (Cat. No. 450030)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli cells Invitrogen™

Zero Blunt® TOPO® PCR Cloning Kit provides a highly efficient, 5-minute, one-step cloning strategy ("TOPO® Cloning") for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. Each kit has the Zero Blunt® TOPO® vector (see figure) containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. Zero® Blunt® TOPO® kits are available with a variety of competent cells, or without competent cells, depending on your needs and budget. Features of the Zero® Blunt® TOPO® PCR Cloning Kits:

Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations
Simple —no ligase, post-PCR procedures, or PCR primers containing specific sequences are required.

Zero Blunt® TOPO® Cloning Kits—overview

Vector: pCR™Blunt II-TOPO® vector—contains the ccdB gene, allowing for direct selection of recombinants

Cloning method: TOPO® cloning—Topoisomerase I–based, 5-minute ligation of blunt-ended PCR products amplified with proofreading thermostable polymerases

Competent cells: Various options—choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells, or use your own.

pCR™Blunt II-TOPO® vector—designed for proofreading polymerases
The pCR™Blunt II-TOPO® vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum® Pfx DNA Polymerase. The pCR™Blunt II-TOPO® vector contains:

• EcoRI sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and Zeocin™ resistance genes for your choice of selection in E. coli
• SP6 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening

pCR™Blunt II-TOPO® clone selection
pCR™-Blunt II-TOPO® allows for direct selection of recombinants via disruption of the lethal E. coli gene, ccdB. The vector contains the ccdB gene fused to the C-terminus of the LacZα fragment. Ligation of a blunt-end PCR product disrupts expression of the LacZα-ccdB gene fusion, permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white screening is not required.

Simplified TOPO®-based cloning
Using TOPO® cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform E. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones, saving you time and money. The pCR™Blunt II-TOPO® vector used in this kit comes with lethal ccdB gene for efficient selection of blunt-ended PCR recombinants products generated bythermostable proofreading polymerases.

The standard in cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

Zero Blunt® TOPO® PCR Cloning® Kit options
The Zero Blunt® TOPO® PCR Cloning Kit for direct insertion of blunt-ended PCR products into a plasmid vector can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

General cloning: TOP10 cells (Cat. No. K2800-20, K2800-40)
High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K2860-01, K2860-40)
General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2820-20, K2820-40)
Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K2830-20)
Provide your own: for flexibility and to save money (Cat. No. 450254)

We also offer a version of the kit that includes a PureLink® Quick Plasmid Miniprep Kit (Cat. No. K2800-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

T4 DNA Polymerase Invitrogen™

T4 DNA Polymerase is a DNA polymerase that has a 3´-exodeoxyribonuclease activity, but lacks 5´a3´ exodeoxyribonuclease activity. A T4 DNA Polymerase Technical Bulletin is available.

Applications:
Labeling double-stranded linear DNA by replacement synthesis (1). Oligonucleotide-directed, site-specific mutagenesis (2). 3´ end-labeling of double-stranded DNA (3). Polishing both 5´ or 3 overhangs to make blunt ends (4).

Source:
Purified from E. coli expressing the T4 DNA Polymerase gene on a plasmid.

Performance and Quality Testing:
Single- and double-stranded endodeoxyribonuclease and phosphatase assays; exodeoxyribonuclease and polymerase activities tested.

Unit Definition:
One unit incorporates 10 nmol of total deoxyribonucleotide into acid-precipitable material in 30 min. at 37°C using DNase I-nicked DNA as template•primer.

Unit Reaction Conditions:
50 mM glycine-NaOH (pH 8.8), 16.6 mM (NH4)2SO4 , 6 mM MgCl2 , 6.5 µM EDTA, 10 mM 2-mercaptoethanol, 0.165 mg/ml BSA, 1.6 mg/ml DNase I-nicked salmon testes DNA, 0.33 mM dCTP, 0.33 mM dATP, 0.33 mM dGTP, 0.33 mM dTTP, 76 nM [3H]dTTP, and enzyme in 0.1 ml for 30 min. at 37°C.

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Electrocomp™ E. coli Invitrogen™

Zero Blunt® TOPO® PCR Cloning Kits offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. The kits include linearized and topoisomerase I-activated pCR™-Blunt II-TOPO® Vector for 5-minute benchtop ligations without ligase (1). The pCR™-Blunt II-TOPO® Vector also features:

ccdB gene for positive selection (2)
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and Zeocin™ resistance genes for your choice of selection in E. coli
• SP6 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening

Selected Zero Blunt® TOPO® PCR Cloning Kits are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

WT Expression Kit, high throughput Invitrogen™

The WT Expression Kit is an RNA amplification kit that is designed to generate amplified sense-strand cDNA ready for fragmentation and labeling using the Affymetrix® GeneChip® WT Terminal Labeling Kit.

Features of the WT Expression Kit:

• Optimized for use with Affymetrix® GeneChip® Human, Mouse, and Rat 1.0 ST Arrays
• Allows RNA samples as small as 50 ng to be analyzed on Affymetrix® GeneChip® Arrays
• Supports a streamlined workflow that does not require a separate rRNA depletion step
• Is ideal for high-sensitivity expression profiling

Eliminates separate rRNA depletion step, preserves transcriptome coverage

The first-generation Affymetrix® amplification method requires depletion of ribosomal RNA (rRNA) from RNA samples for optimal exon-level analysis. Conversely, the Ambion® WT Expression Kit uses a novel reverse transcription (RT) priming method that eliminates the need for a separate rRNA depletion step. The kit includes RT primers designed using a proprietary oligonucleotide matching algorithm that eliminates primer sequences with homology to known ribosomal RNAs. The result is complete and unbiased coverage of the transcriptome, with rRNA amplification levels significantly lower than those of other methods.

Consistent results from less input RNA
With the Ambion® WT Expression Kit, samples as small as 50 ng of total RNA can be analyzed on Affymetrix® GeneChip® Human, Mouse, and Rat Exon and Gene 1.0 ST Arrays. Previous methods required 1 µg of total RNA—often difficult or impossible to obtain from limited sources such as stem cells or small tissue samples. The modest requirement for input RNA permits the analysis of rare samples and provides for more efficient and cost-effective experimentation for most sample types. To demonstrate the performance of the WT Expression Kit, RNA from three sample types (HeLa cells, and Microarray Quality Control (MAQC) A and B samples) was prepared in triplicate using either the Affymetrix® GeneChip® WT cDNA Synthesis and Amplification Kit or the WT Expression Kit, and analyzed on Human Exon 1.0 ST Arrays. Total RNA (1 µg) prepared by the Affymetrix® protocol underwent an rRNA-depletion step while just 50 ng total RNA (20-fold less) was prepared for microarray analysis using the WT Expression Kit. Both sets of samples were handled according to the manufacturers' recommendations. Direct correlation of log2 ratios for MAQC samples A and B was high, (r >0.94) at both the exon and transcript levels. Correlation of log2 ratios with gene expression data obtained using TaqMan® Gene Expression assays for real-time PCR was also high for both kits (see Product Bulletin for data).

Identify more differential expression with less RNA
In addition to its lower input RNA requirement, the WT Expression Kit provides a significant increase in sensitivity. A greater number of probe sets detected above background was obtained at the exon level with the WT Expression Kit as a result of an increased signal-to-noise ratio. A comparison of differential expression analysis results from the two methods indicated that, while differentially expressed genes and exons of MAQC A and MAQC B samples were similar between the two kits (78% and 89%, respectively), significantly more differential expression was observed on arrays hybridized with samples prepared using the WT Expression Kit. 499 more genes and 6,850 more exons were found to be differentially expressed in samples prepared using the Ambion® kit, compared to samples prepared with the Affymetrix® kit.

Note: For high-throughput robotics.

Yeast tRNA Invitrogen™

Yeast tRNA is suitable for use as a carrier in nucleic acid purification and precipitation procedures. It is supplied in lyophilized form.

TOPO™ TA Cloning™ Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli (Supply Center Packaging) Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction, and are available with a variety of competent cells, or no competent cells, depending on your needs and budget. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice. This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

Features of TOPO® TA Cloning® Kits for Sequencing:

Get more sequence—allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations

TOPO® TA Cloning® Kits for Sequencing—overview

Vector: pCR™4-TOPO® TA vector—optimized cloning vector for improved sequencing results

Cloning method: TOPO® TA Cloning®—Topoisomerase I–based, 5-minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector

Competent cells: Various options—choose from kits with general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells, or use your own

pCR™4-TOPO® TA vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA clone selection and manipulation
The pCR™4-TOPO® TA vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection and blue/white screening. The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.

Simplified TOPO®-based cloning
Using TOPO® cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform with the providedE. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones and save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products, which contain 3´A overhangs.

The standard in cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

TOPO® TA Cloning® Kits for Sequencing—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)
• Provide your own: for flexibility and to save money (Cat. No. 450030)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

Mouse Cot-1 DNA Invitrogen™

Mouse Cot-1 DNA® is mouse DNA that is predominantly 50 to 300 bp in size and enriched for repetitive DNA sequences such as the B1, B2, and L1 family members. Mouse Cot-1 DNA® is commonly used to block non-specific hybridization in microarray screening. It can also be used to suppress hybridization of rodent repetitive DNA sequences when mapping rodent clones by in situ hybridization or Southern blotting and to identify mouse clones in screens of somatic-cell hybrid libraries derived from mouse-hamster hybrid cells (1-3). The amount supplied is sufficient for 5 to 10 Southerns or 500 in situ hybridizations.

Performance and Quality Testing:
Purity and DNA size are verified by agarose gel electrophoresis. Concentration is verified spectrophotometrically by diluting Mouse Cot-1 DNA® 1:100 in 50 mM NaOH and using the conversion factor 0.033 µg/µl/A 260 . Other methods used to determine concentration may yield varying results.

Gal-Screen™ β-Galactosidase Reporter Gene Assay System for Yeast or Mammalian Cells Invitrogen™

The Gal-Screen® assay system combines direct cell lysis with rapid, ultra-sensitive chemiluminescent detection of β-galactosidase reporter enzyme.

• Homogeneous assay format allows detection of β-galactosidase in the presence of normal culture media without removal of media and without an additional cell lysis step, providing the easiest and most streamlined assay procedure possible.
• The kinetics of the glow-assay provides a "window" during which measurements may be performed, facilitating HTS applications where assay automation is used.
• Can be used with either mammalian or yeast model systems, providing flexibility in choice of model systems.
• Wide dynamic range of β-galactosidase assay enables accurate measurement of enzyme from femtogram to nanogram range.
• Assay sensitivity is 100 to 1,000-fold better than either the isotopic⁄non-isotopic assays for chloramphenicol acetyl transferase (CAT) or the colorimetric⁄fluorescent assays for β-galactosidase, providing greater sensitivity than competing assay technologies.
• Highly sensitive assay with a wide dynamic range permits detection of high and low levels of reporter without performing numerous sample dilutions.
• Non-radioactive reporter gene assay kit eliminates concerns over use of radioisotopes.
• Assay can be completed in about one hour, providing fast assay turnaround.

Ideal for Screening
This homogeneous assay is ideally suited for screening applications where assay automation is required. The Gal-Screen® system uses Galacton-Star® chemiluminescent substrate for convenient measurement in a luminometer. Light emission reaches maximum in 60-90 minutes and remains constant for 45-90 minutes.

For Research Use Only. Not for use in diagnostics procedures.

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ MAX Efficiency DH5α-T1R E. coli Invitrogen™

Zero Blunt® TOPO® PCR Cloning Kits offers the fastest and easiest method for high-efficiency (cloning of blunt-end PCR products amplified with proofreading thermostable polymerases. The kits include linearized and topoisomerase I-activated pCR™-Blunt II-TOPO® Vector for 5-minute benchtop ligations without ligase (1). The pCR™-Blunt II-TOPO® Vector also features:

ccdB gene for positive selection (2)
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and Zeocin™ resistance genes for your choice of selection in E. coli
• SP6 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening

Selected Zero Blunt® TOPO® PCR Cloning Kits are available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli Invitrogen™

The Zero Blunt® TOPO® PCR Cloning Kit for Sequencing provides a highly efficient, 5-minute, one-step cloning strategy for the direct insertion of proofreading-polymerase–amplified, blunt-ended PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4Blunt-TOPO® vector (see figure) with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice.

pCR™4Blunt-TOPO® vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4Blunt-TOPO® vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4Blunt TOPO® vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4Blunt-TOPO® clone selection and manipulation
The pCR™4Blunt-TOPO® vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection. The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.

Simplified, TOPO®-based cloning
Using TOPO® cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform the provided E. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, which saves you time and money. The pCR™4Blunt-TOPO® vector used in this kit comes with no overhangs for efficient ligation of PCR products created by proofreading, thermostable polymerases that leave blunt-ended PCR products.

The standard in cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

Zero Blunt® TOPO® PCR Cloning Kit for Sequencing—kit formats
The Zero Blunt® TOPO® PCR Cloning Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending on what your needs are:

• General cloning: TOP10 cells (Cat. No. K2875-J10, K2875-20, K2875-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K2880-20, K2880-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2895-20)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K2835-20)

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TaqMan™ Protein Assays Buffer Kit Applied Biosystems™

TaqMan® Protein Assays Buffer Kit contains the buffers required for preparing assay probes and use them in TaqMan Protein Assays experiments. The kit contains dilution buffers and a storage buffer, all of which are used in combination with the TaqMan® Protein Assays Oligo Probe Kit.(4448549)

Human Cot-1 DNA™ Invitrogen™

Human Cot-1 DNA® is commonly used to block nonspecific hybridization in microarray screening. It can also be used to suppress repetitive DNA sequences for the direct mapping of human DNA or mapping genomic clones to panels of somatic-cell hybrids for chromosome localization by Southern blotting. Human Cot-1 DNA® is effective as a library-screening probe for somatic-cell hybrid libraries and flow-sorted chromosome libraries made from somatic-cell hybrids.

About Cot-1 DNA®
Human Cot-1 DNA® is placental DNA that is predominantly 50 to 300 bp in size and enriched for repetitive DNA sequences such as the Alu and Kpn family members. The amount supplied per package is sufficient for 5–10 Southern or 500 in situ hybridizations.

Performance and quality testing
Purity and DNA size are verified by agarose gel electrophoresis. Concentration is verified spectrophotometrically by diluting Human Cot-1 DNA® 1:100 in 50 mM NaOH and using the conversion factor 0.033 µg/µL/A260. Other methods used to determine concentration may yield varying results.

Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing, with One Shot™ TOP10 Electrocomp™ E. coli Invitrogen™

The Zero Blunt® TOPO® PCR Cloning Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of proofreading-polymerase–amplified blunt-end PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4Blunt-TOPO® vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice.

Get More Sequence—Allows for more insert sequence and less vector sequence when using standard sequencing primers

Fast and Easy—Go from PCR-to-clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—Achieve up to 95% clones with correct insert
Proven—Reliable performance for over a decade with over 4000 citations

Zero Blunt® TOPO® Cloning Kits for Sequencing Kit Overview
VECTOR: pCR™4Blunt-TOPO® Vector—Optimized cloning vector for improved sequencing results
CLONING METHOD: TOPO® Cloning —Topoisomerase I based 5 minute ligation of proofreading-polymerase-amplified PCR products with blunt ends to the vector
COMPETENT CELLS: Various Options —Choose from kits with either general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells

pCR™4Blunt-TOPO® VectorOptimized for Sequencing
We have removed much of the multiple cloning site from the pCR™4Blunt-TOPO® vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4Blunt TOPO® vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4Blunt-TOPO® Clone Selection and Manipulation
The pCR™4Blunt-TOPO® vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection. The vector’s minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.

Simplified TOPO®-Based Cloning
Using TOPO® cloning technology there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform the provided E. coli competent cells.

Efficient Cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones to help you save time and money. The pCR™4Blunt-TOPO® vector used in this kit comes with no overhangs for efficient ligation of PCR products created by proofreading, thermostable polymerases that leave blunt-ended PCR products.

The Standard in Cloning
When it comes to cloning, TOPO® cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO® cloning has been applied to many different vectors for a wide array of applications.

Zero Blunt® TOPO® PCR Cloning Kits for Sequencing Kit Options
The Zero Blunt® TOPO® PCR Cloning Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending on what your needs are:

• General cloning: TOP10 (Cat. No. K2875-J10, K2875-20, K2875-40)
• High efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K2880-20, K2880-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2895-20)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K2835-20)

TOPO® products are For Research Use Only. Not intended for any animal or human therapeutic of diagnostic use.

Related Links
Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells Invitrogen™

TOPO® TA Cloning® Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include the pCR™II-TOPO® TA vector with dual T7 and SP6 promoters. By eliminating time-consuming and tedious restriction site cloning, TOPO® cloning is the most reliable cloning method, featuring a 3-step protocol and 5-minute cloning reaction, and yielding up to 95% recombinants.

Convenient features of this TOPO® TA Cloning® Dual Promoter kit include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot® TOP10 Competent cells for added convenience and highest cloning efficiencies

Kit Options: TOPO® TA Cloning® Dual Promoter Kits
TOPO® TA Cloning® Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4600-J10, K4600-01, K4600-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K4660-01, K4660-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4620-01, K4620-40)
• Fast growth: Mach1™ -T1R chemically competent E. coli (Cat. No. K4610-20)
• Repressor/induction needs: TOP10F’ cells (Cat. Nos. K4650-01, K4650-40)
• Provide your own cells (Cat. Nos. 451641, 450641)
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