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| 产品编号 | 产品名称 | 产品包装 | 产品价格 |
| R7035S | RNA 5’ Pyrophosphohydrolase (RppH) | 200U | 468.00元 |
| R7035M | RNA 5’ Pyrophosphohydrolase (RppH) | 1kU | 1758.00元 |
碧云天生产的RNA 5’ Pyrophosphohydrolase (RppH)，即RNA 5’焦磷酸水解酶，是由碧云天自主研发的PerfectProtein™技术平台通过大肠杆菌表达、纯化获得的可用于催化5’端三磷酸化RNA (ppp-RNA)水解去除其无机焦磷酸(Inorganic pyrophosphate, PPi)而获得5’端单磷酸化RNA (p-RNA)的重组酶[1]。RppH蛋白也可以将二腺苷五磷酸(Ap5A)分解成ADP和ATP [2]。RppH发挥活性需要Mg2+ [3]，其催化的反应如下所示。
ppp-RNA → p-RNA + PPi
本产品可以用于RNA的5’末端分析，5’端三磷酸化RNA配合XRN-1的剔除，以及5’端三磷酸化小RNA去焦磷酸后建库测序等。
RppH，又称NudH/YgdP，属于Nudix水解酶超家族。在细菌中，它通过控制数百个mRNA的降解而影响着细菌毒力、应激反应和DNA修复等[3]。在大肠杆菌体内，经RppH催化产生的5’端单磷酸化mRNA可被RNase E所降解[4]。在体外，经RppH催化产生的5’端单磷酸化RNA可被5’-phosphate-dependent exonuclease (XRN-1)水解为单个的核糖核苷酸[5]。
碧云天RNA 5’ Pyrophosphohydrolase (RppH)对5’端三磷酸化RNA 5’端无机焦磷酸的水解效果请参考图1。
图1. 碧云天RNA 5’ Pyrophosphohydrolase (RppH) (R7035)与国外N公司(Competitor N)的RppH对5’端三磷酸化RNA 5’端无机焦磷酸的水解效果对比图。首先，通过体外转录制备300-mer ppp-RNA底物：转录模板质粒线性化并回收，以线性化的质粒为模板，使用转录试剂盒T7 Quick High Yield RNA Transcription Kit (R7016)进行体外转录，并纯化转录产物。测定300-mer ppp-RNA浓度并分装，-80ºC保存。将碧云天与N公司的RppH (5U/µl)分别稀释1.25、2.5、5、10、20、40倍，然后配制10µl反应体系：10mM Tris-HCl (pH7.9), 50mM NaCl, 10mM MgCl2, 1mM DTT, 20U RNase Inhibitor (R0102), 0.5μg 300-mer ppp-RNA, 2U XRN-1 (R7040), 1μl上述不同稀释度的RppH或Storage (Dilution) Buffer，37ºC孵育30分钟，随后95ºC加热5分钟。加热结束后，每个样品中加入2µl DNA上样缓冲液(6X) (D0071)，进行1.5%琼脂糖凝胶电泳，全部上样，160V电泳6分钟。碧云天的RppH与N公司的产品相比，对5’端三磷酸化RNA 5’端无机焦磷酸具有基本一致的水解效果。图中数据仅供参考，不同实验条件下获得的数据会有一定的差异。
来源：由大肠杆菌表达，表达基因为大肠杆菌的RppH基因。
分子量：约19.7kDa。
活性定义：One unit is the amount of enzyme that converts 1µg 300 mer RNA transcript into a XRN-1 digestible RNA in 30 minutes at 37ºC。
纯度：不含DNA内切酶和外切酶，不含RNA酶。
Storage (Dilution) Buffer: 20mM Tris-HCl (pH7.5), 200mM NaCl, 0.1mM EDTA, 1mM DTT, 0.01% Triton X-100 and 50% Glycerol.
10X Reaction Buffer: 100mM Tris-HCl (pH7.9), 500mM NaCl, 100mM MgCl2, 10mM DTT.
失活或抑制：氟离子会对RppH的活性造成抑制。此外，核苷酸和警报酮(Alarmones)也会抑制RppH的活性[4]。
包装清单：| 产品编号 | 产品名称 | 包装 |
| R7035S-1 | RNA 5’ Pyrophosphohydrolase (RppH) (5U/μl) | 40μl |
| R7035S-2 | 10X Reaction Buffer | 1ml |
| R7035S-3 | Storage (Dilution) Buffer | 1ml |
| - | 说明书 | 1份 |
| 产品编号 | 产品名称 | 包装 |
| R7035M-1 | RNA 5’ Pyrophosphohydrolase (RppH) (5U/μl) | 200μl |
| R7035M-2 | 10X Reaction Buffer | 5ml |
| R7035M-3 | Storage (Dilution) Buffer | 5ml |
| - | 说明书 | 1份 |
-20ºC保存，至少两年有效。
注意事项：酶使用时宜存放在冰盒内或冰浴上，使用完毕后宜立即放置于-20ºC保存。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
| Dilution Ratio | RppH Concentration |
| 1.25 | 4U/μl |
| 2.5 | 2U/μl |
| 5 | 1U/μl |
| 10 | 0.5U/μl |
| 20 | 0.25U/μl |
| 40 | 0.125U/μl |
| Reagent | Control | Sample | ||
| Volume | Conc. or amount | Volume | Conc. or amount | |
| Ultrapure Water | 4.5μl | - | 4.5μl | - |
| 10X Reaction Buffer | 1μl | 1X | 1μl | 1X |
| RNase Inhibitor (40U/μl) | 0.5μl | 20U | 0.5μl | 20U |
| pppRNA (0.5μg/μl) | 1μl | 0.5μg | 1μl | 0.5μg |
| Storage (Dilution) Buffer | 1μl | - | - | - |
| RppH (Diluted) | - | - | 1μl | 0.125-4U |
| XRN-1 (1U/μl) | 2μl | 2U | 2μl | 2U |
| Total volume | 10μl | - | 10μl | - |
| 产品编号 | 产品名称 | 包装 |
| D2308-10μg | pT3/SP6/T7-RNA Linearized Template (RNA体外转录质粒) | 10μg |
| D2310-1μg | pT3/SP6/T7-RNA-Template (RNA体外转录质粒) | 1μg |
| D2310-100μg | pT3/SP6/T7-RNA-Template (RNA体外转录质粒) | 100μg |
| D2312-1μg | pRNA-T3-T7 (RNA体外转录质粒) | 1μg |
| D2312-100μg | pRNA-T3-T7 (RNA体外转录质粒) | 100μg |
| D2314-1μg | pRNA-SP6-T7 (RNA体外转录质粒) | 1μg |
| D2314-100μg | pRNA-SP6-T7 (RNA体外转录质粒) | 100μg |
| D6128S | BsaI | 1kU |
| D6128M | BsaI | 5kU |
| D6128L | BsaI | 20kU |
| D6128XL | BsaI | 200kU |
| D7383-1ml | NTP set (100mM each, Nuclease free) | 4×250μl |
| D7385-500μl | NTP Mix (10mM each, Nuclease free) | 500μl |
| D7385-2ml | NTP Mix (10mM each, Nuclease free) | 2ml |
| D7387-250μl | NTP Mix (25mM each, Nuclease free) | 250μl |
| D7387-1ml | NTP Mix (25mM each, Nuclease free) | 1ml |
| R0021 | DEPC水(DNase、RNase free) | 100ml |
| R0022 | DEPC水(DNase、RNase free) | 500ml |
| R0081-1ml | BeyoMag™ RNA Clean Magnetic Beads (RNA纯化磁珠) | 1ml |
| R0081-5ml | BeyoMag™ RNA Clean Magnetic Beads (RNA纯化磁珠) | 5ml |
| R0081-20ml | BeyoMag™ RNA Clean Magnetic Beads (RNA纯化磁珠) | 20ml |
| R0081-100ml | BeyoMag™ RNA Clean Magnetic Beads (RNA纯化磁珠) | 100ml |
| R0102-2kU | RNase Inhibitor | 2000U |
| R0102-10kU | RNase Inhibitor | 10000U |
| R0102-50kU | RNase Inhibitor | 50000U |
| R0107 | 氧钒核糖核苷复合物(RNase抑制剂) | 2ml |
| R0108 | 氧钒核糖核苷复合物(RNase抑制剂) | 10ml |
| R0123 | RNase and DNase Away | 250ml |
| R0125 | RNase, DNase and DNA Away | 250ml |
| R0127 | RNase, DNase, RNA and DNA Away | 250ml |
| R7006S | SP6 RNA Polymerase | 1KU |
| R7006M | SP6 RNA Polymerase | 5KU |
| R7006L | SP6 RNA Polymerase | 25KU |
| R7006XL | SP6 RNA Polymerase | 100KU |
| R7009S | T3 RNA Polymerase | 1KU |
| R7009M | T3 RNA Polymerase | 5KU |
| R7009L | T3 RNA Polymerase | 25KU |
| R7009XL | T3 RNA Polymerase | 100KU |
| R7012S | T7 RNA Polymerase | 1KU |
| R7012M | T7 RNA Polymerase | 5KU |
| R7012L | T7 RNA Polymerase | 25KU |
| R7012XL | T7 RNA Polymerase | 100KU |
| R7016S | T7 Quick High Yield RNA Transcription Kit | 25次 |
| R7016M | T7 Quick High Yield RNA Transcription Kit | 100次 |
| R7018S | T7 High Yield RNA Transcription Kit | 25次 |
| R7018M | T7 High Yield RNA Transcription Kit | 100次 |
| R7020S | SP6 High Yield RNA Transcription Kit | 25次 |
| R7020M | SP6 High Yield RNA Transcription Kit | 100次 |
| R7023S | Pyrophosphatase, Inorganic (E. coli) | 10U |
| R7023M | Pyrophosphatase, Inorganic (E. coli) | 50U |
| R7023L | Pyrophosphatase, Inorganic (E. coli) | 200U |
| R7023XL | Pyrophosphatase, Inorganic (E. coli) | 1000U |
| R7025S | Pyrophosphatase, Inorganic (yeast) | 10U |
| R7025M | Pyrophosphatase, Inorganic (yeast) | 50U |
| R7025L | Pyrophosphatase, Inorganic (yeast) | 200U |
| R7025XL | Pyrophosphatase, Inorganic (yeast) | 1000U |
| R7027S | Thermostable Inorganic Pyrophosphatase | 250U |
| R7027M | Thermostable Inorganic Pyrophosphatase | 1250U |
| R7027L | Thermostable Inorganic Pyrophosphatase | 5KU |
| R7027XL | Thermostable Inorganic Pyrophosphatase | 25KU |
| R7040S | XRN-1 | 20U |
| R7040L | XRN-1 | 100U |
| R7070S | E. coli Poly(A) Polymerase | 100U |
| R7070M | E. coli Poly(A) Polymerase | 500U |
| R7070L | E. coli Poly(A) Polymerase | 2.5kU |
| R7070XL | E. coli Poly(A) Polymerase | 10kU |
| R7075S | Poly(A) Polymerase Tailing Kit | 50-250次 |
| R7090FT | Thermostable RNase H | 50U |
| R7090S | Thermostable RNase H | 250U |
| R7090M | Thermostable RNase H | 1000U |
| R7090L | Thermostable RNase H | 5000U |
| R0722S | Circular RNA Synthesis Kit | 20次 |
| R0722M | Circular RNA Synthesis Kit | 100次 |
| ST876-500ml | BeyoPure™ Ultrapure Water (DNase/RNase-Free, Sterile) | 500ml |
| ST876-100ml | BeyoPure™ Ultrapure Water (DNase/RNase-Free, Sterile) | 100ml |
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