The AMPD1 product, DNase I component D5307, is not tested for protein content, so it is not possible to determine the activity units per mg of protein. This variability in active protein content within a preparation makes it more accurate to sell the product based on its unit activity. If the protocol specifies a certain number of milligrams of DNaseI, it is advisable to determine the specific activity of the enzyme used in the procedure. Alternatively, a titration analysis may be necessary to determine the appropriate amount to use.
AMPD1
DNase I
Amplification Grade
Synonym(s):
Deoxyribonuclease I
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About This Item
Pricing and availability is not currently available.
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Quality Level
form
liquid
concentration
1 unit/μL
technique(s)
RT-PCR: suitable
color
colorless
shipped in
wet ice
storage temp.
−20°C
Related Categories
General description
Deoxyribonuclease I (DNase I) is an endonuclease isolated from bovine pancreas that digests double and single stranded DNA into oligo and mononucleotides. Amplification Grade DNase I has been purified to remove RNase activity, and is suitable for eliminating DNA from RNA preparations prior to sensitive applications, such as RT-PCR (Reverse Transcriptase - Polymerase Chain Reaction).
DNase I digests double- and single-stranded DNA into oligo- and mononucleotides. Using the Reaction Buffer provided, DNA is removed from RNA preparations in a 15 minute digestion at room temperature. The DNase I is then inactivated by heating with the Stop Solution. Heating also denatures hairpins in the RNA, so the RNA can be used directly in reverse transcription.
No current RNA isolation procedure removes 100% of the DNA. Many commercial DNase I formulations are contaminated with residual RNases. This RNase contamination can destroy or degrade valuable RNA samples prior to reverse transcription. Laboratory comparisons have shown that Sigma′s Amplification Grade DNase I demonstrates lower RNase activity than that from several leading molecular biology product suppliers.
DNase I digests double- and single-stranded DNA into oligo- and mononucleotides. Using the Reaction Buffer provided, DNA is removed from RNA preparations in a 15 minute digestion at room temperature. The DNase I is then inactivated by heating with the Stop Solution. Heating also denatures hairpins in the RNA, so the RNA can be used directly in reverse transcription.
No current RNA isolation procedure removes 100% of the DNA. Many commercial DNase I formulations are contaminated with residual RNases. This RNase contamination can destroy or degrade valuable RNA samples prior to reverse transcription. Laboratory comparisons have shown that Sigma′s Amplification Grade DNase I demonstrates lower RNase activity than that from several leading molecular biology product suppliers.
Application
Amplification grade DNase I has been used:
- for the digestion of DNA during isolation and purification of RNA. The purified RNA can be used for the synthesis of cDNA using RNA reverse transcriptase.[1][2][3][4]
- to hydrolyze extracellular matrix (ECM) components and enhance photosensitizer penetration into the biofilm to determine the efficacy of antimicrobial photodynamic therapy (aPDT) on Candida albicans biofilms[5]
- to remove contaminating DNA from total RNA extracted from cattle blood samples[6]
Features and Benefits
- Suitable for the elimination of DNA from RNA
- Minimal RNase activity available
- Optimized 10× reaction buffer and Stop Solution for complete inactivation of DNase I
Suitability
Suitable for use in removing DNA from RNA preparations.
Unit Definition
One unit completely digests 1 μg of plasmid DNA to oligonucleotides in 10 min. at 37 °C.
Legal Information
Purchase of this product is accompanied by a limited license for use in the Polymerase Chain Reaction (PCR) process for research purposes only and in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by an up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., and authorized thermal cycler.
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Storage Class Code
10 - Combustible liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
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Protocols
The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms.
Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.
The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms.
The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms.
Related Content
The Spectrum Plant Total RNA Kit incorporates a new, patent-pending lysis and binding chemistry. This kit is intended to purify total RNA from species and tissue where most Total RNA protocols fail. Without sacrificing affordability, you can purify more total RNA per isolation.
Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.
Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.
Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.
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How can I convert 'units' to mass when adding DNaseI to a buffer solution?
1 answer-
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What is the concentration of the DNAse in Product AMPD1, DNase I?
1 answer-
The DNAse is provided at 1 unit/uL in 1 mL total volume.
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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What do I do if my DNA is not completely digested after 15 minute digestion at room temperature when using Product AMPD1, DNase I?
1 answer-
If further digestion is required, incubation can be performed for 30 minutes at 37°C.
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