Gotaq long pcr master mix kit

Based on information from the Bionano analysis and the Snap Gene program (GSL Biotech LLC, San Diego, CA, USA), a map of the predicted deletions and translocations was generated. PrimerBlast was used to select suitable forward and reverse primers for each aberration (see Table S3) [22 (link)]. Long-distance touchdown PCRs [23 (link)] were performed using the GoTaq^®Long PCR Master Mix Kit (Promega GmbH, Walldorf, Germany) following manufacturer recommendations. The expected PCR products with a length of 5 to 15 kbp were separated on a 0.8% agarose gel applying a 2.6 V/cm distance between the electrodes. Primer walking was used to amplify the breakpoint. The identified breakpoints were re-confirmed by a breakpoint-spanning PCR using the ALLin™ HS Red Taq Master Mix Kit (highQu GmbH, Kraichtal, Germany). Primers and annealing temperatures are listed in Table S3. The expected PCR products had a length of 600 to 1000 bp and were separated on a 2% agarose gel applying 8 V/cm distance between the electrodes. All PCR products were sequenced at a commercial lab (Microsynth Seqlab GmbH, Göttingen, Germany).