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  • ParkerKB
    replied
    Originally posted by LFMar View Post
    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.

    Leave a comment:


  • vidushi
    replied
    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?

    Leave a comment:


  • vidushi
    replied
    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..

    Leave a comment:


  • LFMar
    replied
    Originally posted by vidushi View Post
    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.
    Attached Files

    Leave a comment:


  • vidushi
    replied
    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]

    Leave a comment:


  • LFMar
    replied
    Thanks RCJK and Eric.

    Leave a comment:


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

    Leave a comment:


  • LFMar
    replied
    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.

    Leave a comment:


  • oyjenvol
    replied
    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]

    Leave a comment:


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

    thx

    Leave a comment:


  • giakr84
    replied
    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

    Leave a comment:


  • Apollo704
    replied
    MVE Protocols?

    Does anyone have MVE protocols?

    Leave a comment:


  • jade
    replied
    cDNA sequencing

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

    Leave a comment:


  • mccallumj
    replied
    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.

    Leave a comment:


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

    Leave a comment:

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