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pLenti6.3/V5-DEST™ Gateway™ Vector Kit - FAQs

查看更多产品信息 pLenti6.3/V5-DEST™ Gateway™ Vector Kit - FAQs (V53306)

14 个常见问题解答

可以使用BP Clonase酶和LR Clonase酶替代BP Clonase II 酶LR Clonase II酶进行BP/LR Clonase反应的一步法实验方案吗？

在BP/LR Clonase反应的一步法实验方案中，不建议用BP Clonase酶和LR Clonase酶替代BP Clonase II 酶/LR Clonase II酶，因为这样的重组效率非常低。

有推荐的一步式BP/LR重组实验方案吗？

有的，我们能提供针对BP/LR Clonase反应的一步式实验方案DNA可以在一步反应后被克隆到目的载体中，从而节省了您的时间和金钱。

如果丢失了入门克隆，如何将目的基因从一个Gateway兼容的表达克隆转移到一个新的目的载体？

建议使用一个供体载体进行一次BP反应以获得一个入门克隆。然后将这一入门克隆和目的载体进行一次LR反应以获得新的表达克隆。

我可以单独购买5X LR Clonase缓冲液或5X BP Clonase缓冲液吗？

5X LR Clonase缓冲液或5X BP Clonase缓冲液不作为单独产品出售。它们作为酶试剂盒的一部分进行销售。

是否提供用于在植物内表达的Gateway载体吗？

我们不提供任何用于在植物内表达的Gateway载体。

Can I perform the single-step protocol for the BP/LR Clonase reaction using BP Clonase enzyme and LR Clonase enzyme instead of BP Clonase II enzyme and LR Clonase II enzyme?

In the single-step protocol for the BP/LR Clonase reaction, we would not recommend substituting the BP Clonase II/LR Clonase II enzymes with BP Clonase /LR Clonase enzymes as this would result in very low recombination efficiency.

Do you have a recommended single-step protocol for BP/LR recombination?

Yes, we have come up with a single-step protocol for BP/LR Clonase reaction (http://www.thermofisher.com/us/en/home/life-science/cloning/gateway-cloning.html#1), where DNA fragments can be cloned into Destination vectors in a single step reaction, allowing you to save time and money.

How can I move my gene of interest from a Gateway-adapted expression clone to a new Destination vector as I have lost the entry clone?

We would recommend performing a BP reaction with a Donor vector in order to obtain an entry clone. This entry clone can then be used in an LR reaction with the Destination vector to obtain the new expression clone.

Can I purchase the 5X LR Clonase buffer or 5X BP Clonase buffer separately?

We do not offer the 5X LR Clonase buffer and 5X BP Clonase buffer as standalone products. They are available as part of the enzyme kits.

Do you offer Gateway vectors for expression in plants?

We do not offer any Gateway vectors for expression in plants.

I sequenced one of your vectors after PCR amplification and observed a difference from what is provided online (or in the manual). Should I be concerned?

Our vectors have not been completely sequenced. Your sequence data may differ when compared to what is provided. Known mutations that do not affect the function of the vector are annotated in public databases.

Are your vectors routinely sequenced?

No, our vectors are not routinely sequenced. Quality control and release criteria utilize other methods.

How was the reference sequence for your vectors created?

Sequences provided for our vectors have been compiled from information in sequence databases, published sequences, and other sources.

Do you offer pLenti6.3/V5-DEST Gateway vector as a stand-alone product?

Unfortunately, we do not offer the pLenti6.3/V5-DEST Gateway vector as a stand-alone product. The pLenti6.3/V5-DEST Gateway vector is offered as a component of the pLenti6.3/V5-DEST Gateway Vector Kit (Cat. No. V53306) or as part of the ViraPower HiPerform Lentiviral Gateway Expression Kit (Cat. No. K5330-00).

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