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SuperScript™ II Reverse Transcriptase
For superior cDNA synthesis performance upgrade to Invitrogen™ SuperScript™ IV Reverse Transcriptase.
SuperScript™ II Reverse Transcriptase
Invitrogen™

SuperScript™ II Reverse Transcriptase

Invitrogen SuperScript II Reverse Transcriptase is a genetically engineered MMLV reverse transcriptase (RT) with reduced RNase H activity and increasedRead more
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Catalog NumberNo. of Reactions
18064071200 Reactions
1806402210 Reactions
1806401450 Reactions
Catalog number 18064071
Price (USD)
1,583.65
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1,818.00
Save 234.35 (13%)
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No. of Reactions:
200 Reactions
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Price (USD)
1,583.65
Online Exclusive
1,818.00
Save 234.35 (13%)
Each
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Invitrogen SuperScript II Reverse Transcriptase is a genetically engineered MMLV reverse transcriptase (RT) with reduced RNase H activity and increased thermal stability compared to wild-type MMLV RT. Mutations in the RNase H domain of the enzyme eliminate degradation of the RNA during first-strand cDNA synthesis, which results in higher yields of full-length cDNA.

SuperScript reverse transcriptases are the most highly trusted and widely used RTs with over 50,000 citations, reviews, and publications to date. The enzyme is purified to near homogeneity from E. coli containing the modified pol gene of Moloney Murine Leukemia Virus.

Features of SuperScript II Reverse Transcriptase include:
• High cDNA yields and more full-length product
• Generation of cDNA up to 12.3 kb
• First-strand cDNA synthesis at higher temperatures than conventional MMLV RT

Note: The latest member of the SuperScript RT family, SuperScript IV Reverse Transcriptase, features enhanced thermostability, processivity, yields, and performance with any RNA samples, including those of suboptimal purity or integrity.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Concentration200 U/μL
Final Product TypeFirst-Strand cDNA
FormatStand-alone Enzyme
No. of Reactions200 Reactions
Optimal Reaction Temperature42°C
Quantity4 x 10,000 units
Reaction FormatSeparate components
Reagent TypeReverse Transcription
Reverse TranscriptaseSuperScript II
Ribonuclease H ActivityReduced
Shipping ConditionWet Ice
Size (Final Product)Up to 12.3 kb
Starting MaterialRNA
TechniqueReverse Transcription
GC-Rich PCR PerformanceHigh
Reaction Speed50 min.
Unit SizeEach
Contents & Storage

• SuperScript II Reverse Transcriptase, 4 x 50 μL (4 x 10,000 units at 200 U/μL)
• 5X First-strand buffer
• DTT (100 mM)

Store at –20°C.

Have questions about this product? Ask our AI assisted search.
What is the shipping condition for SuperScript II Reverse Transcriptase and SuperScript First-Strand Synthesis System for RT-PCR?
How can I tell if my RT-PCR product is RNA specific?
What is the highest temperature that SuperScript III , SuperScript II, MMLV, or ThermoScript can be used?
Is it necessary to digest first-strand cDNA made with SuperScript III or II Reverse Transcriptase (RT) or ThermoScript RT with RNase H?
Can I use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase (Cat. No. 28025013, 28025021)? Can other reverse transcriptases, such as SuperScript reverse transcriptase, be used in the same way?
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Frequently asked questions (FAQs)

How can I remove genomic DNA contamination from my sample prior to performing RT-PCR?

If amplification products are generated in the control tube/well that contains no reverse transcriptase (i.e., the no-RT control), it may be necessary to eliminate residual genomic DNA from the RNA sample. Use the following protocol to remove genomic DNA from the total RNA preparation.Random primers are the best choice for degraded RNA, RNA with heavy secondary structure, non-polyadenylated RNA, or prokaryotic RNA. It is recommended only for two-step RT-PCR, and typically gives the highest yields, although the cDNA may not necessarily be full length. Oligo(dT) primers are good to use when trying to recover full-length cDNA from 2-step RT-PCR. The reaction is influenced by secondary structure and RNA quality. Gene specific primers should be used for very specific, mainly one-step RT-PCR reactions. Random primers are the best choice for degraded RNA, RNA with heavy secondary structure, non-polyadenylated RNA, or prokaryotic RNA. It is recommended only for two-step RT-PCR, and typically gives the highest yields, although the cDNA may not necessarily be full length. Oligo(dT) primers are good to use when trying to recover full-length cDNA from 2-step RT-PCR. The reaction is influenced by secondary structure and RNA quality. Gene specific primers should be used for very specific, mainly one-step RT-PCR reactions.

Add the following to an autoclaved 0.5 mL microcentrifuge tube on ice:
1.Total RNA, ideally, less than or equal to 1 µg. (See Note 1 below.)
2.1.0 µL of 10X DNase buffer (200 mM Tris, pH 8.3, 500 mM KCl, 20 mM MgCl2).
3.0.1 U-3.0 U of DNase I (RNase-free, Cat. No. 18047019) or 1.0 U Dnase I, Amplification Grade (Cat. No. 18068015. (See Note 2 below.)
4.Bring volume up to 10 µL with DEPC-treated water.
5.Incubate at room temperature for 15 min. (See Note 3 below.)
6.Terminate the reaction by adding 1 µL 25 mM EDTA and heat 10 min at 65 degrees C. (See Note 4 below.)
7.Place on ice for 1 minute.
8.Collect by brief centrifugation. This mixture can be used directly for reverse transcription.

Please note the following:
1.To work with higher quantities of RNA, scale up the entire reaction linearly. Do not exceed 2 µg RNA in the 10 µL reaction. More RNA will increase the viscosity of the solution and prevent the DNAse I from diffusing and finding the DNA.
2.DNAse I, Amplification Grade has been extensively purified to remove trace ribonuclease activities commonly associated with other "RNAse-free" enzyme preparations and does not require the addition of placental RNAse inhibitor.
3.It is important not to exceed the 15 minute incubation time or the room temperature incubation. Higher temperatures and longer times could lead to Mg2+-dependent hydrolysis of the RNA.
4.This procedure requires careful pipetting of all solutions so that the concentration of divalent metal cation (Mg2+) is controlled.
5.Because the DNAse I must be heated to 65 degrees C to inactivate the enzyme, the concentration of free divalent metal ions must be low enough (less than 1 mM) after addition of the EDTA to prevent chemical hydrolysis of the RNA. See references below.
After the addition of EDTA, there is an approximately 1:1 molar ratio of Mg2+ :EDTA. EDTA chelates Mg2+ molecules on a 1:1 molar basis. Therefore, this RNA can be directly used in a reverse transcription reaction. First-strand reverse transcription buffers typically result in a final concentration of 2.5 mM Mg2+. If the reverse transcription buffer does not contain MgCl2, add it to the reaction at a final concentration of 2.5 mM. This results in a net final concentration of approximately 2.25 to 2.5 mM MgCl2.

References on RNA hydrolysis:
Molekulyarnaya Biologiya (1987) 21:1235-1241.
References on the mechanism of hydrolysis by other cations:
Eichorn GL and Butzov JY (1965) Biopolymers 3:79.
Butzov JY and Eichorn GL (1965) Biopolymers 3:95.
Farkas WR (1968) Biochim Biophys Acta 155:401.
The authors of the first paper express the opinion that the mechanism of the nonspecific hydrolysis by cations which proceeds through 2',3' cyclic phosphate formation is similar to that of specific hydrolysis such as RNA splicing.

How much RNA should be employed for first-strand cDNA synthesis?

The amount of RNA template for a cDNA synthesis is highly flexible and depends upon the amount of sample available and an individual's need. In general, 1 µg total RNA is used in a typical 20-µL RT reaction.

Find additional tips, troubleshooting help, and resources within ourReverse Transcription and RACE Support Center.

Should I treat the cDNA with RNase H prior to downstream processing?

Some feel that the RNA in the RNA:DNA duplex after reverse transcription will inhibit PCR primers from annealing and amplifying the cDNA. The RNA is still present when using RNase H-mutant RTs. RNase H frees the cDNA from the RNA. On the other hand, some feel that the 95 degrees C denaturing step will cause the RNA primers to fall off the DNA and therefore RNase H treatment is not necessary. Therefore, this step is optional. For cloning of larger fragments, RNase H treatment can be beneficial.

What percentage of RNA is converted to cDNA when performing reverse transcription?

This depends highly on the quality of the sample. mRNA itself makes up 1-5% of total RNA. Depending on the primer and enzyme used, reverse transcription can covert >70% of that into cDNA.

Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.

I'm setting up my RT reaction and am trying to decide whether I should use random primers, oligo(dT) primer, gene-specific primer, or oligo(dT)/random mix primers. What would you suggest?

Random primers are the best choice for degraded RNA, RNA with heavy secondary structure, non-polyadenylated RNA, or prokaryotic RNA. It is recommended only for two-step RT-PCR, and typically gives the highest yields, although the cDNA may not necessarily be full length. Oligo(dT) primers are good to use when trying to recover full-length cDNA from 2-step RT-PCR. The reaction is influenced by secondary structure and RNA quality. Gene specific primers should be used for very specific, mainly one-step RT-PCR reactions.

Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.

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