EasyScript cDNA Synthesis Kit

Application:

• First strand cDNA synthesis for PCR
• Construction of cDNA libraries
• Generation of probes for hybridization

EasyScript cDNA Synthesis Kit is a complete system for the efficient synthesis of first strand cDNA from RNA templates. The recombinant RNasin Ribonuclease Inhibitor, supplied with the kit effectively protects RNA template from degradation.

The kit is also supplied with both oligo(dT) and random primers. The oligo(dT) anneals selectively on the poly(A) tail of mRNA. Random primers do not require the presence of poly(A). Therefore, they can be used for transcription of the 5'-end regions of mRNA. Gene-specific primers may also be used with the kit. The first strand of cDNA can be directly used as a template in PCR.

EasyScript Reverse Transcriptase (RTase) within the kit is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript and EasyScript Plus are RNase H deficient (negative). The enzyme is purified from bacteria host as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components:

Storage Buffer:
50mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage: 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol:

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended. 
1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes.
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes.
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes:
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product.
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a random primer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.

EasyScript Reverse Transcriptase

Application:

• Synthesis cDNA froma single-stranded RNA or DNA primer extension
• Sequencing dsDNA
• cDNA library
• Template production for use in PCR
• 3'-end labeling of duplex DNA via end-filling reactions

EasyScript Reverse Transcriptase is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript and EasyScript Plus are RNase H deficient (negative). The enzyme is purified from bacteria h o s t as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components:

Storage Buffer:
50mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage: 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol:

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended.

1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes. 
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes. 
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes:
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product. 
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a randomprimer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.

Model D4 Hippo Electrophoresis System

Gel Size: 16cmW x 17cmL
Footprint: 27cmW x 23cmL x 14cmH
Running Buffer Volume: 1.8L
Optimal Flexibility

The Owl Hippo Horizontal Agarose Gel Electrophoresis System is designed to provide flat, even banding patterns and consistent results with hassle-free gel casting. No tape, grease, agarose seals or other accessories are required. The model D4's design allows you to run one gel tray (51 samples) or stack two gel trays (102 samples) in the buffer chamber while saving valuable bench space. Each of the (2) U.V. Transmissible (UVT) gel trays, 16cmWx 17cmL, accommodates up to (3) combs, allowing the user to run up to 3 series of samples of equal distances. A stand-alone casting platform is included for casting 2 gels simultaneously. A single gel can be cast right in the buffer chamber. Custom combs are available upon request.

Complete System Includes:

• Buffer Chamber
  
• Three Point Leveling Base
  
• SuperSafe Lid with Attached Power Supply Leads
  
• 2 UVT Gel Trays
  
• 6 Combs: 1.5mm Thick
  
• External Gel Caster
  
• Casting Tray
• 3 year warranty

Model D3-14 Centipede Wide Gel Electrophoresis System

Gel Size: 23cmW x 14cmL
Footprint: 27cmW x 30cmL x 11cmH
Running Buffer Volume: 800ml
Great for Screening PCR Samples

The Owl Centipede horizontal model D3-14 wide gel system offers a simple, convenient and fast method for screening multiple samples on a single agarose gel. Ideal for screening PCR samples, plasmid preps, restriction mapping and cloning. 25 to 200 samples can be run simultaneously on one gel; producing clear, tight banding patterns with no "smiling". 

The Centipede System incorporates Owl's EasyCast casting design with an external caster. The UVT gel tray is fitted with a gasket which allows the tray to fit snugly into the external gel caster to provide a leakproof seal without tape. Sample loading is greatly enhanced using micro well pipette format combs (and a multichannel pipette) engineered specifically for use with this system. Using these combs and a multichannel pipette, samples may be loaded directly from a 96-well plate, 8 or 12 at a time. Sample loading with a multichannel pipette speeds up transfer and reduces the chance of sample order error. The unique electrode placement allows this gel to be run quickly without compromising the resolution.

• Buffer Chamber
  
• SuperSafe Lid with Attached Power Supply Leads
  
• EasyCast Gasketed U.V. Transmissible (UVT) Gel Tray
  
• External Gel Caster
  
• 4 Combs: 50 Well, 1.5mm Thick
  
• Bubble Level
• 3 year warranty

Model D2 Spider Wide Gel Electrophoresis System

Gel Size: 14.4cmW x 10.2cmL
Footprint: 20cmW x 19cmL x 10cmH
Running Buffer Volume: 600ml
Great for Protocols with a Large Number of Samples

This horizontal wide gel system can run from 10 to 80 samples on one gel. The wide gel size allows you to load 15 or 30 samples per row with a multichannel pipette format combs. This economical unit has multi functions: it is great to screen PCR samples or use for routine DNA fragment analysis. The small footprint allows it to fit on the most cluttered lab bench.

This system incorporates Owl's EasyCast casting design with an external caster. The UVT gel tray is fitted with a gasket which allows the tray to fit snugly into the external gel caster to provide a leakproof seal without tape. Sample loading is greatly enhanced using micro well pipette format combs engineered specifically for use with this system. Using these combs and a multichannel pipette, samples may be loaded directly from a 96-well plate, 8 or 12 at a time. This speeds up transfer and reduces the chance of sample order error. The unique electrode placement allows this gel to be run quickly without compromising the resolution.

Complete System Includes:

• Buffer Chamber
  
• SuperSafe Lid with Attached Power Supply Leads
  
• EasyCast Gasketed U.V. Transmissible (UVT) Gel Tray
  
• External Multiple Gel Caster
  
• 4 Combs: (2) 30 Well and (2) 40 Well, 1.5mm Thick
• 3 year warranty

PR1MA™ Mini Gel Electrophoresis System

• Complete system, includes gel box, power supply and casting sets
• Quick and easy gel casting, no tape, clamps or spacers required
• Compact design, footprint: 7 x 5.1 in.

The PR1MA™ Mini Gel electrophoresis system includes all of the equipment that you need to get up and running: gel tank, power supply and two casting sets. Ideal for personal use, small laboratories or the classroom, this gel box/power supply combination is compact and easy to use.

The mini gel box attaches directly to the power supply for a secure connection every time. Selectable parameters include 3 preset voltages – 35V, 50V or 100V – and a 99-minute timer. All are set using the membrane keypad.

The safety lid is vented to help dissipate heat generated during electrophoresis and features a viewing area above the gel. Current will only run through the system when the lid is properly in place on top of the gel box.

Trays and casting stands for both 10.5 x 6 cm and 5 x 6 cm gels are included with the MyGel Mini system, as are double-sided combs. The smaller tooth combs are compatible with multi-channel pipettes. Casting is quick and easy – just place the tray in the casting stand and pour. The trays feature a dark contrast strip to help visualize wells while loading and lines that fluoresce under UV and blue light to track progress of the run.

Specifications

ENDURO VE20 Vertical Gel Electrophoresis System

Enduro VE20 Vertical Electrophoresis System (20 x 20 cm), includes glass plates, 2 x 24 well combs, cooling coil and casting base

• Large format vertical gel system
• Electroblotting and gel electrophoresis in same universal gel tank
• Quick assembly system for ease of use
• Complete system allows casting of gels and running of gels at same time increasing efficiency

The ENDURO VE20 is Labnet's new large format vertical gel electrophoresis system. It distinguishes itself in its flexibility of applications and ease of gel casting and set-up requiring only 4 screws to secure 4 gels. The same gel tank can house a large format electroblotting assembly sold separately. This system is ideal for first- and second dimension SDS-PAGE, native, preparative, gradient and high-resolution nucleic acid electrophoresis.

Innovative screw-clamp technology consists of only four screws to secure up to four 20 x 20cm gels. This vertical screw clamp technology distributes pressure evenly along the height of gel rather than in the center to eliminate plate bowing and gel compression, while maintaining a leak-proof seal during casting.

NO TANK TOP ASSEMBLY
A built-in inner buffer chamber within the PAGE insert allows set up to be completed without inclusion of an upper buffer chamber.

CASTING ADVANTAGES

• Dual purpose PAGE insert eliminates time consuming transfer of glass plates between separate casting and gel running components.
• Glass plates with bonded spacers assure clean well formation, correct alignment for leak-free casting and eliminates manually aligning spacers.
• Casting stand design ensures leak-free casting even if glass plates are misaligned.

Specifications:

Screen up to 7 Samples on 1 Gel in Less than 5 Minutes

The Owl Pollywog horizontal micro gel electrophoresis system is designed for quick restriction fragment analysis. Glass slides, 2" x 3", are used as the gel tray. Thin gels can be poured and formed by surface tension directly on the glass slide. Because the gel is so thin, a complete gel run will last 5 to 10 minutes or gels can be cast and solidified in less than 10 minutes using the available caster (see below). This system is designed for quick rapid screening and can be used to check a few PCR samples before you load a large number of samples on a larger gel, saving time.

Complete System Includes:

• Buffer chamber
• Lid
• Power supply leads
• Gel casting tray
• Casting slides (10)
• 1 comb, 7 well, 1.5 mm
• 3 year warranty

Gel Size: 12cmW x 14cmL
Footprint: 18cmW x 24.5cmL x 9.5cmH
Running Buffer Volume: 800ml

The Owl EasyCast horizontal B2 mini gel system provides the most flexibility in a very small footprint. The longer gel length and the two comb slots give you the flexibility of running 8 to 48 samples on one gel. The optional multi load tray can increase the maximum number of samples to 108 in a multichannel pipette format. The simple to use casting method, and multiple comb and tray options, makes this Owl.s most popular system.

Owl offers a wide range of combs. The two comb slots on the U.V. Transmissible (UVT) gel tray, double your sample capacity by doubling the number of sample wells. Heavy duty, one piece combs eliminate comb assembly. UVT gel trays are silk screened with a florescent ruler for easy measurement of bands. This system is available with optional buffer exchange ports, catalog number B2-BP, for use with an external recirculation pump.

Complete System Includes:

Model B1 EasyCast Mini Gel Electrophoresis System

Gel Size: 9cmW x 11cmL
Footprint: 15cmW x 22cmL x 9.5cmH
Running Buffer Volume: 600ml
Fits Most Teaching Applications

The Owl EasyCast model B1 horizontal mini gel system is the perfect small device. Run from 5 to 34 samples in this functional system. The unit is economical enough for every researcher to have one at their workstation. The durability and ease of use makes this the perfect choice for students and high volume researchers.

Owl offers a wide range of combs. The two comb slots on the U.V. transmissible (UVT) gel tray, double your sample capacity by doubling the number of sample wells. Heavy duty, one piece combs eliminate comb assembly. UVT gel trays are silk screened with a florescent ruler for easy measurement of bands. This system is available with optional buffer exchange ports, catalog number B1-BP, for use with an external recirculation pump.

Complete System Includes:

• Buffer Chamber
  
• SuperSafe Lid with Attached Power Supply Leads
  
• EasyCast Gasketed U.V. Transmissible (UVT) Gel Tray
  
• 2 Combs: 10 & 14 Well, Double Sided, 1.0/1.5mm Thick
• 3 year warranty

Model B1A EasyCast Mini Gel Electrophoresis System

Gel Size: 7cmW x 8 cmL
Footprint: 10.5cmW x 16 cmL x 9.5 cmH
Running Buffer Volume: 400ml 
Ideal Size for Every Lab

The EasyCast horizontal mini gel system is designed to provide flat, even banding patterns and consistent results with less hassle. This is the perfect size to determine molecular weight ranges of PCR fragments or larger sized DNA molecules. The all in one design allows you the option of casting and running a gel in the buffer chamber, eliminating the need for additional equipment. Run from 5 to 24 samples in 15 minutes.

The two comb slots on the U.V. Transmissible (UVT) gel tray, double your sample capacity by doubling the number of sample wells. Heavy duty, one piece combs eliminate comb assembly. UVT gel trays are silk screened with a florescent ruler for easy measurement of bands. This system is available with optional buffer exchange ports, catalog number B1A-BP, for use with an external recirculation pump.

Complete System Includes:

• Buffer Chamber
  
• SuperSafe Lid with Attached Power Supply Leads
  
• EasyCast Gasketed U.V. Transmissible (UVT) Gel Tray
  
• 2 Combs: 6 & 10 Well, Double Sided, 1.0/1.5mm Thick
• 3 year warranty

Model B1 EasyCast Mini Gel Electrophoresis System

Gel Size: 9cmW x 11cmL
Footprint: 15cmW x 22cmL x 9.5cmH
Running Buffer Volume: 600ml
Fits Most Teaching Applications

The Owl EasyCast model B1 horizontal mini gel system is the perfect small device. Run from 5 to 34 samples in this functional system. The unit is economical enough for every researcher to have one at their workstation. The durability and ease of use makes this the perfect choice for students and high volume researchers.

Owl offers a wide range of combs. The two comb slots on the U.V. transmissible (UVT) gel tray, double your sample capacity by doubling the number of sample wells. Heavy duty, one piece combs eliminate comb assembly. UVT gel trays are silk screened with a florescent ruler for easy measurement of bands. This system is available with optional buffer exchange ports, catalog number B1-BP, for use with an external recirculation pump.

Complete System Includes:

• Buffer Chamber
  
• SuperSafe Lid with Attached Power Supply Leads
  
• EasyCast Gasketed U.V. Transmissible (UVT) Gel Tray
  
• 2 Combs: 10 & 14 Well, Double Sided, 1.0/1.5mm Thick
• 3 year warranty