What are the activities and applications of Exonuclease III, Mung Bean Nuclease, and S1 Nuclease?
Exonuclease III catalyzes the removal of mononucleotides from a 3'-OH terminus of duplexed DNA. It requires a substrate of double-stranded DNA containing a blunt end or a 3' recessed end. Exonuclease III also works at nicks to generate gaps.
S1 nuclease is an endonuclease specific for single-stranded DNA or RNA and can be used to study nucleic acid hybridization, mapping RNA start sites and RNA splice sites. This enzyme is five times more active on DNA than RNA, and it will digest all nucleic acids if the enzyme is added to the reaction in excess.
Mung bean nuclease is an endonuclease similar in action to S1 nuclease. Unlike S1 nuclease, it is used to generate blunt ended DNA from ss overhangs.
Answer Id: 2954
What is the maximum thermal output for the Applied Biosystems® 7500 / 7500 Fast Sequence Detection System in BTU/h?
Will dye labeling with the Silencer® siRNA Labeling Kits affect the functionality of the siRNA?
There is always a risk that any modification to a molecule could have some effect on its function, but our R&D scientists have found that dye labeling does not generally affect the activity of siRNA. Also, labeled siRNA appears to have no toxic effect on cells at normal transfection levels.
Answer Id: 2881
How can I tell if my RT-PCR product is RNA specific?
Include a control reaction where the RNA has not been incubated with reverse transcriptase to test for specificity. If this RNA gives a PCR product, it is most likely generated from genomic DNA contamination. Alternatively, a primer set spanning two different exons can be designed such that the PCR product from the cDNA would be of a different size compared to a product generated from genomic DNA. Primers may also be designed to span an exon/exon junctions. These primers are not likely to amplify from genomic DNA templates. For DNase treatment of RNA, we recommend using Amplification-Grade DNase I, Cat. No. 18068-015, or an equivalent product.
Answer Id: 2926