Whole genome amplification (WGA) was performed with REPLI-g kit (Qiagen,
MD), which utilizes the proofreading enzyme Phi 29 polymerase to achieve
high-fidelity amplification of genomic DNA48 (link),49 (link). For sorted
samples with yield cell number larger than 1000, cells were washed with PBS and
then resuspended with 5μl of sterile PBS. REPLI-g mini kit (Qiagen, MD)
was used for WGA according to the manufacturer’s protocol. For sorted
samples with less than 1000 cells or single cell analysis, cells were sorted
into 5μl PBS, thereafter, REPLI-g single cell kit (Qiagen, MD) was used
for WGA according to the manufacturer’s protocol. For DNA samples, we
used 1 to 10ng DNA as input, and REPLI-g mini kit (Qiagen) was used for the WGA.
All the products of WGA were purified with Agencourt AMPure XP beads (Beckman
Coulter, IN) to remove residual dNTP, primers and random products with size
< 100bp.
MD), which utilizes the proofreading enzyme Phi 29 polymerase to achieve
high-fidelity amplification of genomic DNA48 (link),49 (link). For sorted
samples with yield cell number larger than 1000, cells were washed with PBS and
then resuspended with 5μl of sterile PBS. REPLI-g mini kit (Qiagen, MD)
was used for WGA according to the manufacturer’s protocol. For sorted
samples with less than 1000 cells or single cell analysis, cells were sorted
into 5μl PBS, thereafter, REPLI-g single cell kit (Qiagen, MD) was used
for WGA according to the manufacturer’s protocol. For DNA samples, we
used 1 to 10ng DNA as input, and REPLI-g mini kit (Qiagen) was used for the WGA.
All the products of WGA were purified with Agencourt AMPure XP beads (Beckman
Coulter, IN) to remove residual dNTP, primers and random products with size
< 100bp.
