ULYSIS™ Alexa Fluor™ 594 Nucleic Acid Labeling Kit, 588/615 nm - FAQs

查看更多产品信息 ULYSIS™ Alexa Fluor™ Nucleic Acid Labeling Kit - FAQs (U21654, U21652, U21650, U21660)

17 个常见问题解答

ULYSIS标记是否兼容于基因芯片分析？

是的，已经有许多使用ULYSIS标记探针进行基因芯片分析的例子。这里是一些供您参考的文献：

•Babak T, Zhang W, Marros Q et al. (2004) Probing microRNAs with microarrays: tissue specificity and functional inference. RNA 10(11):1813–1819.
•Hiley SL, Jackman J, Babak T et al. (2005) Detection and discovery of RNA modifications using microarrays. Nucleic Acids Res 33(1):e2.
•Torchet C, Badis G, Devaux F et al. (2005) The complete set of H/ACA snoRNAs that guide rRNA pseudouridylations in Saccharomyces cerevisiae. RNA 11(6):928–938.

可以将ULYSIS核酸标记试剂盒标记的探针保存起来以后使用吗？

ULYSIS标记的探针可以在任意缓冲液、TE、福尔马林、杂交缓冲液或乙醇中长期保存。我们建议使用您的常规存储条件保存探针，只需保持避光。ULYSIS偶联物是非常稳定的。避免苯酚。

对于使用ULYSIS核酸标记试剂盒进行RNA标记，你们有什么诀窍吗？

这里是一个从DNA标记实验方案修改而来的初步实验方案：不要使用核酸酶处理RNA，而是直接在90°C孵育10分钟或在85°C孵育1分钟进行标记。每1μg RNA加入2 μg糖原，然后通过乙醇沉淀进行纯化。请参考以下文献：

•Babak T, Zhang W, Marros Q et al. (2004) Probing microRNAs with microarrays: tissue specificity and functional inference. RNA 10(11):1813–1819.
•Hiley SL, Jackman J, Babak T et al. (2005) Detection and discovery of RNA modifications using microarrays. Nucleic Acids Res 33(1):e2.
•Torchet C, Badis G, Devaux F et al. (2005) The complete set of H/ACA snoRNAs that guide rRNA pseudouridylations in Saccharomyces cerevisiae. RNA 11(6):928–938.

ULYSIS试剂盒可以用来标记长于1000碱基对的探针甚至质粒吗？

用ULYSIS核酸标记试剂盒标记更大的探针是可能的，但是可能需要将染料稀释以避免（或至少减少）聚合现象的发生。请参考这篇文献： Coelho-Castelo AA, Santos Júnior RR, Bonato VL et al. (2003) B-lymphocytes in bone marrow or lymph nodes can take up plasmid DNA after intramuscular delivery. Hum Gene Ther 14(13):1279–1285.

ULYSIS标记的DNA在高温时稳定吗？

使用ULYSIS核酸标记试剂盒标记的寡核苷酸在100°C可以稳定存在5分钟，并且在68°C保存过夜不会引起复合物的解离。

在使用ULYSIS核酸标记试剂盒标记和沉淀之后回收的DNA很少。我怎么才能提高回收率呢？

在标记和纯化DNA之后，您可以向其中加入10 µg鲑精DNA。这将提高您沉淀后的回收率。

我可以在柱纯化后直接将ULYSIS核酸标记试剂盒标记的DNA用于杂交反应吗？

我们建议先使用乙醇沉淀将样本浓缩并计算DNA浓度和标记度。

你们可以提供一份对于ULYSIS核酸标记试剂盒的概述吗？

ULYSIS寡核苷酸化学标记试剂可以实现快速简便的将荧光染料加到核酸聚合物的嘌呤碱基上。这一方法又叫做通用连接系统 (Universal Linkage System，ULS)，是基于铂金修饰的染料在鸟嘌呤N7位形成一个稳定的加合物。ULYSIS标记探针可以用于荧光原位杂交（FISH）和许多其它应用。

Is ULYSIS labeling compatible with microarray analysis?

Yes, there are numerous examples of ULYSIS labeled probes that have been used in microarray analysis. Here are a few publications for your reference:

- Babak T, Zhang W, Marros Q et al. (2004) Probing microRNAs with microarrays: tissue specificity and functional inference. RNA 10(11):1813-1819.
- Hiley SL, Jackman J, Babak T et al. (2005) Detection and discovery of RNA modifications using microarrays. Nucleic Acids Res 33(1):e2.
- Torchet C, Badis G, Devaux F et al. (2005) The complete set of H/ACA snoRNAs that guide rRNA pseudouridylations in Saccharomyces cerevisiae. RNA 11(6):928-938.

Can probes labeled with the ULYSIS Nucleic Acid Labeling Kits be stored for later use?

Long-term storage for the ULYSIS labeled probes can be done in just about any kind of buffer, TE, formamide, hybridization buffer, or ethanol. We suggest using your normal storage conditions as long as you protect the probes from light. ULYSIS conjugates are very stable. Avoid phenol.

Do you have any tips on using the ULYSIS Nucleic Acid Labeling Kits for RNA labeling?

A preliminary protocol modifies our DNA-labeling protocol: Do not nuclease-treat the RNA, but label it directly by incubating for 10 minutes at 90°C or 15 minutes at 85°C. Add 2 µg of glycogen for every 1 µg of RNA and purify by ethanol precipitation. Refer to these publications:

- Babak T, Zhang W, Marros Q et al. (2004) Probing microRNAs with microarrays: tissue specificity and functional inference. RNA 10(11):1813-1819.
- Hiley SL, Jackman J, Babak T et al. (2005) Detection and discovery of RNA modifications using microarrays. Nucleic Acids Res 33(1):e2.
- Torchet C, Badis G, Devaux F et al. (2005) The complete set of H/ACA snoRNAs that guide rRNA pseudouridylations in Saccharomyces cerevisiae. RNA 11(6):928-938.

Can the ULYSIS kits be used on probes longer than 1,000 base pairs or even plasmids?

It might be possible to label larger probes with the ULYSIS Nucleic Acid Labeling Kits, but the dye will likely need to be diluted to avoid (or at least reduce) problems with aggregation. Refer to this publication: Coelho-Castelo AA, Santos Junior RR, Bonato VL et al. (2003) B-lymphocytes in bone marrow or lymph nodes can take up plasmid DNA after intramuscular delivery. Hum Gene Ther 14(13):1279-1285.

How stable is the ULYSIS labeled DNA to high temperature?

An oligonucleotide labeled with a ULYSIS Nucleic Acid Labeling Kit should survive 100°C for 5 minutes, and storage at 68°C overnight should also not cause any dissociation of the complex.

My recovery of DNA after labeling with ULYSIS Nucleic Acid Labeling Kit, and precipitation is low. What can I do to improve it?

After labeling and purifying your DNA, you can add 10 µg of salmon sperm DNA. This should improve your recovery after precipitation.

Can I use the DNA labeled using ULYSIS Nucleic Acid Labeling Kit, for hybridization directly after column purification?

We recommend using ethanol precipitation to concentrate the sample first to calculate the DNA concentration and degree of labeling.

Could you provide an overview of the ULYSIS Nucleic Acid Labeling Kits?

The ULYSIS oligonucleotide chemical labeling reagents allow you to rapidly and easily couple our fluorescent dyes to purine bases in nucleic acid polymers. The method, the Universal Linkage System (ULS), is based on the use of a platinum coordination complex that forms a stable adduct at the N7 position of guanine. ULYSIS labeled probes are used in fluorescence in situ hybridization (FISH) and numerous other applications.

What are the ULYSIS labeling kits used for?

ULYSIS labeling kits are used for labeling short, unmodified DNA and RNA strands between ~100-1,000 bp for FISH analysis.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.