4
Library preparation
Approx 65 Mins
Goal: to use the Ligation Sequencing Kit to prepare a Lambda DNA library.
Library preparation using the Ligation Sequencing Kit takes approximately 70 minutes.
The protocol consists of two steps:
- FFPE-repairing the DNA, and preparing the fragment ends for adapter ligation
- Ligation of sequencing adapters
Keep all reagents on ice when not in use.
What you will need
- 0.2 ml thin-walled PCR tubes
- 1.5 ml Eppendorf DNA LoBind tubes
- Control Expansion (EXP-CTL001)
- Freshly prepared 70% ethanol in nuclease-free water
- Hula mixer (gentle rotator mixer)
- Ice bucket with ice
- Ligation Sequencing Kit V14 (SQK-LSK114)
- Magnetic rack
- Microfuge
- NEBNext FFPE Repair Mix (NEB, M6630)
- NEBNext Quick Ligation Module (NEB, E6056)
- NEBNext® Ultra II End Repair / dA-tailing Module (NEB, E7546)
- Nuclease-free water (e.g. ThermoFisher, AM9937)
- P10 pipette and tips
- P100 pipette and tips
- P1000 pipette and tips
- P2 pipette and tips
- P20 pipette and tips
- P200 pipette and tips
- Thermal cycler
- Timer
- Vortex mixer