Preliminary

• prepare SBS sequencing kit Box 1 (4°C) & Box 2 (-20°C);
• thaw in warm water and put on ice: IMX36, FFN36, SMX36 & separately: CMX36;
NB! always keep CMX away from other components, change gloves every time you deal with CMX
• record the lot numbers of reagents in the run list;

Prepare

1. prepare incorporation mix:
   • add to IMX36 tube whole content of

     nick volume reagent FFN36 1.75ml Fluorescent nucleotides mix SDP36 220µl DNA polymerase

   • mix by inverting;
2. mix PR1, PR2, PR3, SMX36 by inverting;
3. centrifuge IMX36 and SMX36: 1min, 1000x g, NT;
4. sign tubes/bottles with reagent position number and record their weights

   position nick reagent #1 IMX Incorporation mix #2 PW1 Deionized water #3 SMX Scanning mix #4 PR1 High salt buffer #5 PR2 Incorporation buffer #6 CMX Cleavage mix #7 PR3 Cleavage buffer

5. separately:
   • mix CMX36 by inverting;
   • centrifuge CMX36: 1min, 1000xg, NT;
   • sign tube, record weight;
   • wash/change gloves;