> For the complete documentation index, see [llms.txt](https://clipforum.flow.bio/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://clipforum.flow.bio/experimental/alternative-cdna-library-prep-protocols.md).

# Alternative cDNA library prep protocols

We’ve described a comparison of the variant options for the library preparation protocols in our [2018 review](https://authors.elsevier.com/a/1WUmG3vVUP5-cM).

## **Kits as an alternative for library prep**

**What is the feasibility of doing RNA library prep using a commercially available kit (NEB Ultra, or Illumina Tru-seq), after RNA extraction following proteinase digestion? In this way, the isolated RNA will be treated as a normal RNA sample and random hexamer primers would be used for reverse-transcription.**

The pro is that you can use an established kit, so possibly less optimisation needed. The cons are several. Some are described in our review (greater specificity with on-bead ligation, the capacity for non-radioactive visualisation in irCLIP, etc). In addition, I’m not aware of a kit that would ligate the adapter to cDNA to allow amplification of truncated cDNAs. So when using kits, data will be similar to HITS-CLIP, and restricted to readthrough cDNAs. Also, CLIP RNAs are often too short to allow priming with random hexamers. So all in all, use of kits may work in some cases, but could lead to biased data with limited resolution.<br>


---

# Agent Instructions
This documentation is published with GitBook. GitBook is the documentation platform designed so that both humans and AI agents can read, navigate, and reason over technical content effectively. Learn more at gitbook.com.

## Querying This Documentation
If you need additional information that is not directly available in this page, you can query the documentation dynamically by asking a question.

Perform an HTTP GET request on the current page URL with the `ask` query parameter, and the optional `goal` query parameter:

```
GET https://clipforum.flow.bio/experimental/alternative-cdna-library-prep-protocols.md?ask=<question>&goal=<endgoal>
```

`ask` is the immediate question: it should be specific, self-contained, and written in natural language.
`goal` is optional and describes the broader end goal you are ultimately trying to accomplish on behalf of the user. GitBook uses it to tailor the answer towards what is most useful for that goal.

The response will contain a direct answer to the question and relevant excerpts and sources from the documentation.

Use this mechanism when the answer is not explicitly present in the current page, you need clarification or additional context, or you want to retrieve related documentation sections.
