FastDigest Eco130I

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	The PCR product was treated at 37°C for 20 min andthen at 65°C for 5 min with the Fast Digest Eco130I (StyI)(Fermentas) enzyme and the expected product sizes weredetermined as 391 and 313 bp, respectively.

Protocol tips
The PCR product was treated at 37°C for 20 min andthen at 65°C for 5 min with the Fast Digest Eco130I (StyI)(Fermentas) enzyme and the expected product sizes weredetermined as 391 and 313 bp, respectively.

Publication protocol

"In the sample taken from the bone marrow, FIP1L1-PDGFRA fusion was examined with two different proto-cols at the RNA level. The reaction protocols were per-formed with the primers and probes that were designed andsynthesized specifically to the transcript and withoutprobes based on the manufacturer’s instructions and wereadapted to the real-time PCR device LightCycler (LC) 2.0.FIP1L1-PDGFRA fusion was amplified using the Light-Cycler FastStart DNA Master SYBR Green I Kit besidesforward primer F7 and reverse primer R14 for the RNAsample. Moreover, the PCR protocol we formed with theprimer and probe series designed specifically to the transcriptwas adapted to LC 2.0 by using the FRET (fluorescenceresonance energy transfer) technique with a pair of fluo-rescence-marked hybridization probes. After the productshad been run in 2 % agarose gel, they were imaged under UVlight by using the Vilber Lourmat device. The real-time PCRresult of the cases are presented in Figs.1and2. The gelimages of the cases are presented in Figs.3and4. Restrictionfragment length polymorphism (RFLP): The RFLP methodwas applied to show that the resulting transcript product wasthe real product of FIP1L1-PDGFRA fusion. With this pur-pose, the PCR product was treated at 37°C for 20 min andthen at 65°C for 5 min with the Fast Digest Eco130I (StyI)(Fermentas) enzyme and the expected product sizes weredetermined as 391 and 313 bp, respectively. The RFLPresult of the cases are shown in Figs.5and6.In conclusion, the fusion transcript was found positive atthe RNA level with both PCR methods in two cases fol-lowed in our center

(14) (PDF) Two cases with hypereosinophilic syndrome shown with real-time PCR and responding well to imatinib treatment. Available from: https://www.researchgate.net/publication/232280826_Two_cases_with_hypereosinophilic_syndrome_shown_with_real-time_PCR_and_responding_well_to_imatinib_treatment#fullTextFileContent [accessed Aug 22 2020]."

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