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Monarch RNA Cleanup Kit (500 μg)

Monarch RNA Cleanup Kit (500 μg) カタログ番号:T2050

カタログ番号

サイズ

濃度

価格

保存温度

T2050S 10 preps ¥10,500 室温
T2050L 100 preps ¥85,500 室温

製品カテゴリ>グループ

  • 核酸精製キット>RNAクリーンアップ

特徴

in vitro 転写反応などのクリーンアップに最適な RNA 精製キット (500 µg)

 

特長:

 ・in vitro 転写反応などの RNA 酵素反応中の RNA クリーンアップが可能

 ・DNase I や Proteinase K 処理した Total RNA の精製も可能

 ・RT-qPCR や NGS ライブラリー調製のための RNA クリーンアップに最適

 ・RNA サイズ : ≧ 25 nt (オプション・プロトコールにより ≧15 ntも可能)
 ・溶出液量 : ≧ 50 ul
 ・一般的な回収率 : 70 - 100 %
 ・純度 : A260/280 > 1.8、A260/230 > 1.8

 

  モナーク RNA クリーンアップ・プロトコール

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The Monarch RNA Cleanup Kit (500 µg) is suitable for cleaning up large quantities (>250 µg) of RNA from in vitro transcription reactions

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1. RNA transcripts of varying sizes (0.6-8 kb) were synthesized using the HiScribe™ T7 Quick High Yield RNA Synthesis Kit (NEB #E2050) using 1.5-1.8 µg of DNA template for two hours at 37°C. 40 µl of each in vitro transcription IVT) reaction was cleaned up using the Monarch RNA Cleanup Kit (500 µg, T2050) and eluted in 200 µl. RNA yields were calculated from the resulting A260, measured using a Nanodrop® spectrophotometer and ranged from 268-425 µg of RNA per IVT reaction.


2. RNA integrity (200 ng/lane) was assessed on a 1% agarose-TBE gel stained with SYBR® Gold. 

 

 

 

The Monarch RNA Cleanup Kit (500 µg) cleans up large-scale in vitro transcription reactions and generates yields consistent with other large-scale cleanup kits  

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0.3 and 1.8 kb fragments were in vitro transcribed at 37°C (overnight and 2 hours, respectively) using the HiScribe™ T7 Quick High Yield RNA Synthesis Kit (NEB #E2050). Following DNase I treatment (4 U DNase I, 37°C, 15 minutes), transcription reactions were pooled and 200 µl cleaned up using either the NEB Monarch RNA Cleanup Kit (500 µg) or the MEGAclear Transcription Clean-Up Kit (Thermo Fisher Scientific). In vitro transcribed RNA was eluted twice with 100 µl of nuclease-free water following a 5-minute on-column incubation (room temperature for Monarch and 65°C for MEGAclear. Recovery of the synthesized RNA transcript was calculated from the resulting A260 using a Trinean Dropsense™ 16. The Monarch RNA Cleanup Kit (500 µg) produces similar RNA yields as the MEGAclear Kit for large-scale in vitro transcription reactions.

 

 

 

RNA recovery is increased by incubating the column with the elution buffer (nuclease-free water) prior to the elution spin

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rRNA (16S and 23S Ribosomal Standard from E.coli, Sigma) was purified using the Monarch RNA Cleanup Kit (500 µg, NEB #T2050). 100 µl of nuclease-free water was added to the column and incubated for either 0,1,3 or 5 minutes at room temperature before spinning to elute the RNA. The percent recovery of RNA was calculated from the resulting A260 as measured using a Trinean Dropsense™ 16. A five-minute incubation period produced the maximum yield.

 

 

キット内容

カラム単品とバッファー単品も販売しています。

Monarch RNA Cleanup Column (500 µg)

Monarch RNA Cleanup Binding Buffer

Monarch RNA Cleanup Wash Buffer

Nuclease-free Water

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