{"product_id":"neb-m0526s","title":"New England Biolabs, M0526S, Sce PUS1","description":"This is an  \u003cb\u003eRelated Categories\u003c\/b\u003e Epitranscriptome Analysis,, RNA Modification  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X NEBuffer™ r1.1 Incubate at 30°C 1X NEBuffer™ r1.1 10 mM Bis-Tris-Propane-HCl 10 mM MgCl2 100 µg\/ml Recombinant Albumin (pH 7 @ 25°C)  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 55°C  \u003cb\u003eFAQ\u003c\/b\u003e Q: What is the effect of Mg2+ concentration on Sce PUS1 activity? A: For certain RNA templates we found that the absence of Mg2+ in the reaction buffer can have a stimulatory effect on Sce PUS1 activity, while it can have an inhibitory effect on others. On other RNA templates, Mg2+ may be required for efficient Sce PUS1 activity. Q: How can Sce PUS1 activity be improved when low concentrations of RNA are used? A: Addition of 10% PEG 300 may improve Sce PUS1 activity, especially when low RNA concentrations are used. Q: What effect does MnCl2 have on Sce PUS1 activity? A: MnCl2 at concentrations \u0026gt; 1 mM has an inhibitory effect on Sce PUS1. Q: What is the effect of NaCl concentration on Sce PUS1 activity? A: Sce PUS1 is sensitive to NaCl; concentrations \u0026gt; 50 mM will inhibit Sce PUS1 activity. Q: Can Sce PUS1 modify more than 1 µg of RNA in one reaction? A: Depending on the substrate, additional Sce PUS1 may be necessary to convert \u0026gt; 1 µg of RNA. However, we recommend setting up multiple reactions containing \u0026lt; 1 µg to ensure greater efficiency. Q: Can I scale up or scale down the reaction volume of the Sce PUS1 reaction? A: We observed a drop in Sce PUS1 activity in reaction volumes smaller or larger than 100 µl. Thus, we recommend a minimum reaction volume of 100 µl, and to using multiple 100 µl reactions if scaling up for additional substrate is needed. Q: Can I shorten or extend the incubation time of Sce PUS1 with target RNA? A: We found that the majority of Sce PUS1 targeted uridines are converted to pseudouridines after 2 hours of incubation. However, preferred target sites may be converted after shorter incubation times. Depending on the substrate, incubation for more than 2 hours can further increase the uridine to pseudouridine conversion ratio. Q: Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers? A: NEB is excited to announce that we have switched BSA-containing reaction buffers (NEBuffer 1.1, 2.1, 3.1 and CutSmart® Buffer) to Recombinant Albumin-containing buffers (NEBuffer r1.1, r2.1, r3.1 and rCutSmart™ Buffer). We are also in the process of moving all restriction enzyme formulations to contain rAlbumin. We feel that moving away from animal-containing products is a step in the right direction and are able to offer this enhancement at the same price. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835531317417,"sku":"M0526S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0526s","provider":"Iright","version":"1.0","type":"link"}