Author: Steven Henikoff; Jorja G. Henikoff
Title: Profiling the epigenome at home Document date: 2020_4_17
ID: mojf4l6b_42
Snippet: . CC-BY 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.15.043083 doi: bioRxiv preprint Cycle 1: 58°C for 5 min (gap filling) Cycle 2: 72°C for 5 min (gap filling) Cycle 3: 98°C for 30 sec Cycle 4: 98°C for 10 sec Cycle 5: 60°C for 10 sec Repeat Cycles 4-5 11 times 72°C for 1 min and hold at 8 o C Tip: To minim.....
Document: . CC-BY 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.15.043083 doi: bioRxiv preprint Cycle 1: 58°C for 5 min (gap filling) Cycle 2: 72°C for 5 min (gap filling) Cycle 3: 98°C for 30 sec Cycle 4: 98°C for 10 sec Cycle 5: 60°C for 10 sec Repeat Cycles 4-5 11 times 72°C for 1 min and hold at 8 o C Tip: To minimize the contribution of large DNA fragments and excess primers, PCR should be performed for no more than 12 cycles, preferably with a 10 s 60-63°C combined annealing/extension step. Tip: The cycle times are based on using a conventional Peltier cycler (e.g., BioRad/MJ PTC200), in which the ramping times (3°C/sec) are sufficient for annealing to occur as the sample cools from 98°C to 60°C. Therefore, the use of a rapid cycler with a higher ramping rate will require either reducing the ramping time or other adjustments to assure annealing.
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