Hello,
I am analysing the cost benefit of doing:
RNA extraction and library preparations myself
Sending RNA samples for sequencing
Sending the entire sample for extraction and RNA preparation
If I have 24 samples, does anyone know how long it will take to get the libraries ready?, I know the extraction and quality control can take a day or so. But I wonder how long it takes to prepare the libraries and the possibility of making a mistake since the kits are so expensive.
Thanks
It depends on the kit, you're using.
If you use NEBNext® Ultra™ II RNA Library Prep Kit for Illumina (E7770, plus E7490, plus E7335 e.g.), it will take you about 8 hours to process 8 samples at once, starting from quality-controlled RNA.
Once you become comfortable with the protocol, you can increase the number of samples and do 16 samples at once, which will take you about 10-11 hours to process using multichannel pipettes. Some people do 24 samples at once.
You don't have to do the entire protocol at one day. There are at least 4 "safe stops" in the protocol, where you can freeze your protocol samples at -20C and proceed whenever you can.
At the end you have to do the library quality control using the Agilent DNA chip for example.
Studying the long protocol and getting familiar with all the reagents of the kit is obviously very important and takes about a day.
If you're starting with 8 samples (or less) and following the the protocol carefully, you should be able to get good libraries.
It also helps a lot, if you have seen somebody preparing cDNA libraries. There are always some helpful tricks on how to do the magnet-based extraction steps for example. But there are probably a plenty of video tutorials on that, too.
I hope, this helps.
It depends a lot on the type of sample and the protocol that you are using.
I do RNA extraction, and 'purification' with a tailored protocol (not just one kit) and cDNA libraries in 4 days total (having three stops for freezing). As Jaroslawna mentioned, then it all goes down to how many samples you can manage at the same time. I would recommend starting with just a few, until you know the protocol.
Good Luck! :)
- Badges
- Science topic
- Similar topics
- Analytical Chemistry
- Extraction