NeoPrep Indices

[RezaFalsafi]

Hello
I got a mRNA kit for NeoPrep, the Illumina automated Library prep system. As you may know it only has 24 unique indices
So i tried to use dilutions of my own illumina compatible indices
I used one index at different dilutions and 50% worked and 50% did not and this is the case for every dilution i made
it is really strange why only half worked
Anyone has done anything the same, or any ideas
By the way, i think the adapters provided in the kit are 6 bp or 7 with the A overhang but my adapters from Bioo are 8 bp
Any ideas or suggestions? I diluted them with Tris HCl pH 8.0
Thanks a lot

[RemitoAmigo]

Hi Reza,

Interesting...I was about to try something as well with the i7s of the Truseq HT kit....I didn't know about the Bioo kits.

Yes, they are 6bp in length.

Were you able to sequence the libraries? From the sounds of it...it looks like you need a minimum concentration for the ligation to be efficient. The ones you seem to have are at 0.6uM.

What dilutions have you tried? Which did work and which did not?

I was thinking of measuring left over indexes from the neoprep kit with my Qbit to determine the approximate concentration from the off the shelf neoprep kit. Then replicating the reaction with our own indexes.

Looking forward to hear from you.

Cheers,

Remito