NEBNext^® Small RNA Library Prep Set for SOLiD™

1. Library Preparation Using NEBNext® Small RNA Library Prep Set for SOLiD™ (E6160)

1. Is Index 1 in the NEBNext Small RNA Library Prep Set for SOLiD kit (E6160) compatible with the NEBNext Small RNA Multiplex (1-16) Prep Set for SOLiD kit (E7450)?
2. If my input is <5ug and therefore use 0.5 ul of  3' SR Adapter 3, SR RT Primer 3 and 5' SR Adapter 3, instead of 1.0 ul, my total volumes for each reaction will be short by 0.5 ul. Should I replace that lost volume with water?
3. Why do I add the SR RT Primer 3 before the 5’ ligation instead of right before the reverse transcription step?
4. If I forget to add the 3’ SR RT Primer 3 before the 5’ ligation can I add it before the reverse transcription step and move forward?
5. Can samples be stored longer than 72 hours at -20°C?
6. How many cycles of PCR should I do?
7. If I accidentally move all 30 ul of the 5’ ligation product forward to the reverse transcription mixture will the reaction still work?

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• NEBNext^® Small RNA Library Prep Set for SOLiD™

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