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.

Gel Cutting Tips, 1.1 x 4mm

• Safely excise bands from gels
• Avoid cross contamination
• One-handed operation for quick and accurate gel cutting
• Safer than using razor blades and won't scratch transilluminators

Bullseye 1Kb DNA Ladder, Logic

• Ready to use
• Contains 16 DNA bands: 100bp-10Kb
• 920ng DNA/6µl/loading
• Easy quantification of DNA fragments
• Stable at room temperature
• Supplied with 6x sample loading buffer

Bullseye 1Kb DNA Ladder, Logic contains 16 DNA bands ranging from 100bp to 10Kb, formulated so that each band contains an amount of DNA that correlates logically to its size, allowing the user to estimate both the size and the quantity of specific fragments at a glance. It is particularly useful for protocols such as probe labeling, DNA sequencing and optimizing insert/vector ratio in ligation reactions, where DNA concentration must be taken into consideration.

Size: 1200µl
Storage: Store at -20°C.
Concentration: 920ng/6µl

Loading Buffer Composition:

10mM Tris-HCl
1mM EDTA (pH 8.0)
0.02% Bromophenol blue
0.02% Xylene cyanol
5% Glycerol

Usage: Add 6µl of Bullseye 1Kb DNA Ladder, Logic directly to wells designated for markers, which will yield the amount as indicated in the picture. You may use more than 6µl of ladder, depending on well size and level of dye used to visualize the bands. Calculate the amount for each band according to your loading volume.

Need a great DNA Ladder at a great price? Try PR1MA!  Click here to order.

• A unique combination of PCR products and a number of proprietary plasmids digested with appropriate restriction enzymes to yield 11 fragments, suitable for use as molecular weight standards for agarose gel electrophoresis.
• The DNA includes fragments ranging from 100-1,500 base pairs. The 500 and 1,500 base pair bands have increased intensity to serve as reference points.
• The approximate mass of DNA in each band is provided (0.5µg a load) for approximating the mass of DNA in comparably intense samples of similar size.

Source:

• PCR products and double-stranded DNA digested with appropriate restriction enzymes, are phenol and equilibrated to 10nM Tris-HCl (pH 8.0) and 10mM EDTA.

Range: 100-1,500 bp (DL1); 250-10,000 bp (DL5)
Number of bands: 11 (DL1); 13 (DL5)
Concentration: 100 µg/mL
Package: 50 µg/500 µL
Recommended Load: 5 µL / well
Contains orange G & xylene cyanol FF as tracking dyes. (DL1)
Contains bromophenol blue & xylene cyanol FF as tracking dyes. (DL5)
Storage:

• Store at 25°C for 6 months
• Store at 4°C for 12 months

Store at -20°C for 24 months

Bullseye 100bp DNA Ladder

• Ready to use
• Contains 11 DNA bands: 100-1500bp.
• Clearly identifiable 500bp band as reference
• 500ng DNA/6µl/loading
• Easy to load
• Stable at room temperature
• Supplied with 6x sample loading buffer

Bullseye 100bp DNA Ladder consists of 11 DNA fragments ranging in size from 100-1500 base pairs (bp). 6µl will yield at least 30ng DNA in any single band. The intensity of the 500bp band has been increased to serve as a reference for easy identification.

Size: 1200µl
Storage: Store at -20°C.
Concentration: 500ng/6µl
Loading Buffer Composition:

10mM Tris-HCl
1mM EDTA (pH 8.0)
0.02% Bromophenol blue
0.02% Xylene cyanol
5% Glycerol

Usage: Add at least 6µl Bullseye 100bp DNA Ladder directly to wells designated for markers. You may need more than 6µl of ladder, depending on well size and level of intensity needed to visualize the bands.

Please give us a call for a sample.

Need great dNTPs at a great price? Try PR1MA!  Click here to order.

• Mix of dATP, dCTP, dGTP, dTTP
• Each nucleotide is at a concentration of 12.5 mM
• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labelling and sequencing processes
• High purity: >98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at -20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases

• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labeling and sequencing processes
• High purity: >98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at -20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases

• Mix of dATP, dCTP, dGTP, dTTP
• Each nucleotide is at a concentration of 100 mM
• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labelling and sequencing processes
• High purity: >98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at -20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases

dNTP SET

(dATP, dCTP, dGTP, dTTP)

100 mM

Store at 20°C For in-vitro laboratory use only

Need great dNTPs at a great price? Try PR1MA!  Click here to order.

• Mix of dATP, dCTP, dGTP, dTTP
• Each nucleotide is at a concentration of 10 mM
• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labelling and sequencing processes
• High purity: >98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at -20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases