[ParkerKB]

Has anyone used adaptors with expired date for 454 rapid library preparation? I've bought the adaptors (up to 10 samples) but I used them for just 2 DNA samples, now I have 8 more samples for library prep and the adaptors date just expired. I don't know if I try to use them or if I buy new ones.

We use a 454 GS Junior and have used expired library prep reagents, and I know of other groups that have used them. We have had problems with library preps both with and without expired reagents so I can't say for sure that it will work, but it is certainly worth a shot. Our technical rep suggested a PCR that amps from the adapters--this will tell you if the adapters have successfully ligated and also to check for the approximate size range of the fragments (since we don't have a bioanalyzer). This would let you know whether it is worth pursuing emPCR rather than going through and not getting any enriched beads (we have found that out the hard way). Let me know if you want the details on the PCR.

[vidushi]

hi

can anyone tell me how much time does it take to analyse data generated by NGS? I am doin PhD, so just curious to know?

[vidushi]

NGS data analysis

Thank u so much for the manual.......

can anyone tell me how much time (approx) does it take to analyse the NGS generated data..

[LFMar]

hi
i also need a copy of the 454 Titanium Sequencing Manual please? could you kindly send it to me. It will be a great help for me.
email: [email protected]

I am sending it attached.

[vidushi]

hi
i also need a copy of the 454 Titanium Sequencing Manual please? could you kindly send it to me. It will be a great help for me.
email: [email protected]

[LFMar]

Thanks RCJK and Eric.

[RCJK]

If they've just recently expired, they'd still work and you'll be okay.

[LFMar]

Expired adaptors

Has anyone used adaptors with expired date for 454 rapid library preparation? I've bought the adaptors (up to 10 samples) but I used them for just 2 DNA samples, now I have 8 more samples for library prep and the adaptors date just expired. I don't know if I try to use them or if I buy new ones.

[oyjenvol]

i will use 454 for transcriptome sequencing of ploids
so if anybody have done such work please give me some suggestions
thank you very much
EMAIL: [email protected]

[lidaof]

is there any protocol for library preparation when transcriptome sequencing using 454?

thx

[giakr84]

About number of samples

Hi all,
I need informations,
What's the maximum number of samples that I could put inside a 454 GS Junior's microtiter plate ?
Thanks

[Apollo704]

MVE Protocols?

Does anyone have MVE protocols?

[jade]

cDNA sequencing

Many thanks to Bertie and mccallumj. The info are indeed very helpful!

[mccallumj]

re Poly A etc

We have had very nice results from Ti sequencing of normalised cDNA prepared using Evrogen Mint + Trimmer kits, using a modified 3' Primer XXXXXXXXXXX(T)4G(T)9C(T)10VN as described by Beldade et al.

The information posted at http://www.cees.uio.no/research/faci...equencing.html was very useful.

[Bertie]

jade, take a look at this - it might help.

http://gsl.cals.ncsu.edu/media/cDNA.pdf