Second-strand cDNA was then synthesized in a solution containing buffer, dNTP, RNaseH and DNA polymerase I and subsequently purified using a QiaQuick PCR extraction kit (Qiagen).
The second-strand cDNA was synthesized using buffer, dNTPs, RNase H and DNA polymerase I. Short fragments were purified using a QiaQuick PCR extraction kit and resolved with EB buffer for end repair then adding poly(A).
Second-strand cDNA was synthesised using buffer, dNTPs, RNase H and DNA polymerase I. Fragments were purified using a QIAquick PCR extraction kit and eluted with EB buffer for end reparation and poly(A) addition.
Polymerase chain reaction products were purified using a QIAquick PCR purification kit (Qiagen) prior to sequencing.
Polymerase chain reaction products were purified using a QIAquick PCR purification kit (QIAGEN, Crawley, UK).
Double-stranded (ds) cDNA synthesis was performed using Phusion polymerase (New England Biolabs, Ipswich, MA) with a hot start of 98° for 30 sec, followed by 18 cycles of 98° for 7 sec, 66° for 20 sec, and 72° for 4 min. The ds-cDNA polymerase chain reaction product was purified using a QIAquick PCR Purification column (QIAGEN).
Second strand cDNA was synthesized using DNA polymerase I and RNaseH, and then purified using a QiaQuick PCR extraction kit (Qiagen, Hilden, Germany).
Once the number of cycles was identified, 16 PCRs (50 µL) were completed using high-fidelity Taq DNA polymerase (Clontech), and the products were cleaned using a QiaQuick column (QIAGEN).
The second-strand cDNA was synthesized using buffer, dNTP/dUTP mix, RNase H, and DNA polymerase I. Short double-strand cDNA fragments were purified using a QIAquick PCR extraction kit and eluted with EB buffer for end repair, with the addition of an 'A' base, and ligated to Illumina sequencing adaptors.
Double-strand cDNAs were purified using a QIAquick PCR Purification Kit (QIAGEN, Germany) and processed for end repair and 3' adenylation using Klenow polymerase.
