2026-05-31T01:18:36Z https://api.figshare.com/v2/oai

oai:figshare.com:article/23661813 2023-07-14T16:59:51Z category_24316 portal_549 item_type_3 month_year_07_2023

10.5522/04/23661813.v1 https://figshare.com/articles/dataset/RP-PCR_Traces/23661813 https://ndownloader.figshare.com/files/41525706 https://ndownloader.figshare.com/files/41525709 https://ndownloader.figshare.com/files/41525712 https://ndownloader.figshare.com/files/41525715 https://ndownloader.figshare.com/files/41525718 https://ndownloader.figshare.com/files/41525721 https://ndownloader.figshare.com/files/41525724 https://ndownloader.figshare.com/files/41525727 https://ndownloader.figshare.com/files/41525730 https://ndownloader.figshare.com/files/41525733 https://ndownloader.figshare.com/files/41525736 https://ndownloader.figshare.com/files/41525739 https://ndownloader.figshare.com/files/41525742 https://ndownloader.figshare.com/files/41525745 https://ndownloader.figshare.com/files/41525748 https://ndownloader.figshare.com/files/41525751 https://ndownloader.figshare.com/files/41525754 https://ndownloader.figshare.com/files/41526021 https://ndownloader.figshare.com/files/41526024 https://ndownloader.figshare.com/files/41526027 https://ndownloader.figshare.com/files/41526030 https://ndownloader.figshare.com/files/41526033 https://ndownloader.figshare.com/files/41526036 https://ndownloader.figshare.com/files/41526039 https://ndownloader.figshare.com/files/41526042 https://ndownloader.figshare.com/files/41526045 https://ndownloader.figshare.com/files/41526048 https://ndownloader.figshare.com/files/41526051 https://ndownloader.figshare.com/files/41526054 https://ndownloader.figshare.com/files/41526057 https://ndownloader.figshare.com/files/41526060 https://ndownloader.figshare.com/files/41526063 https://ndownloader.figshare.com/files/41526066 https://ndownloader.figshare.com/files/41526069 https://ndownloader.figshare.com/files/41526270 https://ndownloader.figshare.com/files/41526273 https://ndownloader.figshare.com/files/41526276 https://ndownloader.figshare.com/files/41526279 https://ndownloader.figshare.com/files/41526282 https://ndownloader.figshare.com/files/41526285 https://ndownloader.figshare.com/files/41526288 https://ndownloader.figshare.com/files/41526291 https://ndownloader.figshare.com/files/41526294 https://ndownloader.figshare.com/files/41526297 https://ndownloader.figshare.com/files/41526300 https://ndownloader.figshare.com/files/41526303 https://ndownloader.figshare.com/files/41526306 https://ndownloader.figshare.com/files/41526309 https://ndownloader.figshare.com/files/41526312 https://ndownloader.figshare.com/files/41526315 https://ndownloader.figshare.com/files/41526318 https://ndownloader.figshare.com/files/41526321 https://ndownloader.figshare.com/files/41526324 https://ndownloader.figshare.com/files/41526327 https://ndownloader.figshare.com/files/41526330 https://ndownloader.figshare.com/files/41526333 https://ndownloader.figshare.com/files/41526336 https://ndownloader.figshare.com/files/41526486 https://ndownloader.figshare.com/files/41526489 https://ndownloader.figshare.com/files/41526492 https://ndownloader.figshare.com/files/41526495 https://ndownloader.figshare.com/files/41526498 https://ndownloader.figshare.com/files/41526501 https://ndownloader.figshare.com/files/41526510 https://ndownloader.figshare.com/files/41526513 https://ndownloader.figshare.com/files/41526516 Rocca, Clarissa Clarissa Rocca Kekenadze, Mariam Mariam Kekenadze Turchetti, Valentina Valentina Turchetti Nagy, Sara Sara Nagy Kvirkvelia, Nana Nana Kvirkvelia Beridze, Maia Maia Beridze Kaiyrzhanov, Rauan Rauan Kaiyrzhanov Houlden, Henry Henry Houlden Neurogenetics RPPCR ABI ALS 2023-07-14 2023-07-14 Dataset 2023 University College London C9orf72 expanions tested by RP-PCR. Traces were generated by running the PCR products through the ABI 3730 machine for capillary electrophoresis.  A PCR Mastermix was prepared by mixing 12.5ul of Amplitaq Gold 360 Master Mix (ThermoFisher), 9.5ul of 5M Betaine (ThermoFisher), 1 ul of 10pmol/ul FAM labelled Forward primer (5'-TGTAAAACGACGGCCAGTCAAGGAGGGAAACAACCGCAGCC-3'), 1 ul of 10pmol/ul Reverse primer (5'-CAGGAAACAGCTATGACC-3'),1ul of 10pmol/ul repeat specific reverse primer (5'-CAGGAAACAGCTATGACCGGGCCCGCCCCGACCACGCCCCGGCCCCGGCCCCGG-3') and 1ul of 100ng/ul DNA. Samples were amplified with an initial heat denaturation of 95°C for 10 minutes, followed by 10 cycles of 95°C for 30 seconds, 58°C for 2 minutes, 72°C for 2 minutes, and then 25 cycles of 95°C for 10 minutes, followed by 10 cycles of 95°C for 30 seconds, 58°C for 2 minutes, 72°C for 2 minutes with a 20seconds increase per cycle. The final extension step was 72°C for 7 minutes. The PCR was run on a 9700 Block, at ramp speed 9600. After PCR, 1ul of the reaction product was addedd to a mix with 9.2ul of Formamide (Roche) and 0.1 ul of GeneScan 500 LIZ Size Standard (ThermoFisher).  

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