DYN-R&D SF Plant DNA extraction: user guide

Content: Buffer A: 1X25ml ready to use Buffer B: 2X40ml ready to use Cat No. DYN-SF-02 Buffer C: 1X40ml ready to use Store at room temperature Description: Step 2: Buffer A The SF plant DNA extraction method was developed to 1) Add 50ul of Buffer A into each well of the 96 well meet the need for highest throughput of plant genotyping. PCR plate. The SF plant DNA extraction kit makes use of 3 ready to 2) Spin down the plate for 2min 4000rpm use buffers. 3) Cover plates with PCR seal. The duration of a 96well sample plate extraction is 2 4) Insert the plate into a thermo block. 0 minutes. 5) Heat samples to 98 c for 90 seconds. Make use of Note: The SF DNA plant extraction is customized for the heated lid. downstream DYN-genotyping application. Important: Make use of regular personal protection Step 3: Buffer B equipment: lab coat, gloves, goggles. 1) Carefully open the PCR plate seal. 2) Add 120ul of Buffer B. Storage: Store at room temperature. Step 4: DNA dilution. No detectable reduction of performance after 6 months storage. 1) Prepare 96 well microtest, round bottom shaped plates: add 75ul of Buffer C into each well. Protocol: 2) Transfer 20ul from each well of the PCR plate from Step 1: Sample collection, preparation. step 1-3 into the Buffer C prepared plate. The SF DNA extraction is suitable for: Step 5: Store samples: Plant leaves - ungrounded. 1) Diluted (blue colored) DNA plates can be stored in Seeds- crushed. 0 4 c up to 10 days. 2) Concentrated DNA, PCR plates can be stored in - 0 1. SF sample preparation plant leaves: 20 c forever. It is possible to repeat the DNA a) Cut a 4-5 mm diameter of the leaf. dilution: step 4, over and over as required. b) Insert leaf cut into the bottom of a 96 well PCR plate. Down stream DNA genotyping: c) Do not ground or crush the leaf cut. a) Add 2ul of diluted (blue) DNA into a ready to use 96 d) Place samples in shade until further process. well DYN genotyping plate. e) Samples can be kept up to 10-14 days in room b) Add 1.2ul of diluted (blue) DNA into a ready to use temp until further process. 384 well DYN genotyping plate. c) Follow DYN ready to use genotyping plate's user 2. SF sample preparation seeds. guide. a) Insert seeds into a 96 well micro-test, round bottom shaped plates. One seed into each well. b) Insert 70ul of ddw into each well. Incubate plate over night for seed softening. For some seeds it may require longer incubation. c) Crush seed with a 96 pin seed crusher. d) Transfer 10ul of crushed seeds in each well into a 96 well PCR plate.