Label IT® 核酸標記試劑 – Label IT® Nucleic Acid Labeling Reagents-Mirus

Label IT^® Nucleic Acid Labeling Kit, Cy^®5貨號MR-MIR3725 (25ug) 

Efficient, direct, non-enzymatic labeling of DNA and RNA

Label IT® 核酸標定技術(Label IT® Nucleic Acid Labeling Technology )通過單一步驟(One-step Chemical Method)，標記螢光或酵素分子至核酸樣品( DNA 或 RNA 內的鹼基)。與其他非放射性標定技術例如隨機引發( random priming)、切口平移(nick translation）不同。Label IT® Nucleic Acid標記試劑為共價鍵連接 (Covalent Mechanism)，無需顯著改變起始核酸(starting nucleic acid)或阻礙下游雜交性能(hybridization performance)。Label IT® 反應不會破壞核酸起始材料，因此 Label IT®技術可廣泛用於體內、體外和分子生物學應用。標定的染劑常見的如CX-羅丹明(CX-rhodamine)、螢光素(fluorescein)、MFP488、地高辛(digoxin)、生物素( biotin)、Cy®3、Cy®5、二硝基苯酚 (dinitrophenol , DNP) 或 TM-羅丹明(TM-rhodamine)，由於 Label IT® 反應不會破壞核酸起始材料，因此 Label IT®技術可廣泛用於體內、體外和分子生物學應用。依標準操作流程，標定密度每 20-60 個鹼基對有一個 Label IT® 標定。該標定密度可應用於大多數實驗; 基因表達或 RNAi 研究的應用。如果需要替代的標定密度(labeling density)，只需增加或減少標定量反應中加入試劑或調整反應孵育時間。準確的標定密度請參閱

• 任何 DNA、RNA 模板皆可適用
• 多種螢光、非螢光標定分子可選。
• 化學標定方式，流程簡單，一小時即可完成標定反應。
• 不會破壞標定對象 (核酸分子) 的結構完整性。
• 標定反應不會影響雜交 (hybridization) 效率。

Each Full Size Kit contains sufficient reagents to label 100 µg nucleic acid.
Each Trial Size Kit contains sufficient reagents to label 25 µg nucleic acid.

Label IT® 核酸標定試劑盒。 Label IT® 化學標定試劑由三個區域組成：標記（螢光團或半抗原）（綠色）、促進與核酸靜電相互作用的連接基團（黃色）以及共價連接 Label IT 的反應性烷基化基團（藍色） ® 試劑可與核酸內的任何反應性雜原子結合。 Label IT® 試劑與核酸的連接不會改變核酸的結構或影響下游雜交性能，因此，使用 Label IT® 試劑標記的核酸可用於研究人員定義的多種應用中。

Non-destructive Direct Labeling of Nucleic Acid. 小鼠總 RNA 用 Label IT® Cy®3 (B) 標記，並在 Agilent RNA 6000 Nano 芯片上與未標記的總 RNA (A) 一起進行電泳和分析。對於標定和未標定的樣品，RNA 完整性值 (RIN) 約為 9.0。

Labeling Density can be Easily Controlled with the Label IT® Reagents. Plasmid DNA was labeled with increasing ratios (w:w) of Label IT® CX-Rhodamine to DNA. The extent of labeling was estimated, and normalized to the amount of recovered DNA (# labels/100 base pairs), using two different criteria: fluorescence intensity of the attached rhodamine (excitation at 585 nm, emission at 610 nm), and visible absorbance at 585 nm. The significance of quenching, at the higher labeling ratios, becomes apparent in the fluorimetric estimations of labeling density.

Label IT® MFP488 Labeled Plasmid DNA is Less Sensitive to Photobleaching as Compared to Label IT® Fluorescein Labeled Plasmid DNA. (A) Plasmid DNA was  labeled with Label IT® MFP488 or Label IT® Fluorescein.  Purified DNA was transfected into HeLa cells using the TransIT®-LT1 Transfection Reagent at a 3:1 reagent-to-DNA ratio (vol:wt). Two hours post transfection labeled DNA was visualized using a 63X oil immersion objective on a Zeiss Axiovert Inverted Fluorescence microscope. Images were captured before and after a 2 minute photobleaching period from a FITC light source. (B) The ratio (X-axis) of the brightness of individual puncta was calculated by dividing the photobleached value for fluorescence brightness by the value obtained from non-photobleached values for four separate images using Image J software (NIH), the average and standard deviation are shown. Photobleaching was also measured after 5 minutes for Label IT-MFP488 labeled plasmid DNA and the signal was still 4-fold higher than Label IT® Fluorescein labeled plasmid DNA at 2 minutes (data not shown).

Label IT® MFP488 Labeled Plasmid DNA is More Resistant to the Effects of Low pH as Compared to Label IT® Fluorescein Labeled Plasmid DNA. Plasmid DNA was labeled at a low density, 1 label per 65 bp, using Label IT® MFP488 or Label IT® Fluorescein Nucleic Acid Labeling Kits. Purified DNA was subsequently diluted in various pH buffers (50 mM HEPES, pH 8, 100 mM MES, pH 5.4, and 100 mM MES, pH 3). Fluorescence was measured using a Tecan Genios Fluorometer and normalized to the respective control measured at 50 mM HEPES, pH 8.

Product Name	Pack Size	Cat. No.
Label IT® Nucleic Acid Labeling Kit, Biotin	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3425 MR-MIR3400
Label IT® Nucleic Acid Labeling Kit, CX-Rhodamine	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3125 MR-MIR3100
Label IT® Nucleic Acid Labeling Kit, Cy®3	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3625 MR-MIR3600
Label IT® Nucleic Acid Labeling Kit, Cy®5	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3725 MR-MIR3700
Label IT® Nucleic Acid Labeling Kit, Digoxin	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3325 MR-MIR3300
Label IT® Nucleic Acid Labeling Kit, DNP	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3825 MR-MIR3800
Label IT® Nucleic Acid Labeling Kit, Fluorescein	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR3225 MR-MIR3200
Label IT® Nucleic Acid Labeling Kit, MFP488	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR7125 MR-MIR7100
Label IT® Nucleic Acid Labeling Kit, TM-Rhodamine	labels 25 µg nucleic acid labels 100 µg nucleic acid	MR-MIR4125 MR-MIR4100