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454 Library / Emulsion / Instrument Protocols (October 2008)

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  • #16
    MVE Protocols?

    Does anyone have MVE protocols?

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    • #17
      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

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      • #18
        is there any protocol for library preparation when transcriptome sequencing using 454?

        thx

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        • #19
          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]

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          • #20
            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.

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            • #21
              If they've just recently expired, they'd still work and you'll be okay.

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              • #22
                Thanks RCJK and Eric.

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                • #23
                  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]

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                  • #24
                    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

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                    • #25
                      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..

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                      • #26
                        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?

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                        • #27
                          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.

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