RNase H-dependent PCR enables highly specific amplification of antibody variable domains from single B-cells
Fig 5
Characterization of VL and VH amplicons following amplification with rh-PCR Gen-2 primers.
(A) Agarose gels of standard primers (top row) or rh-PCR Gen-2 primers (bottom row). Gels of the VL fragments are on the left and of the VH fragments on the right. (B) Electropherograms comparing a series of amplicons produced using standard primers (left) to those produced using rh-PCR Gen-2 primers (right). Truncated PCR products (U- Unwanted products) and unused primers (PP- Primer Pool) are denoted by arrowheads. (C) Comparison of wells lacking a full length VL (left) or VH (right) products amplified with either standard or rh-PCR Gen-2 primers. Truncated PCR products and unused primers are captured by a parenthesis and a single arrowhead, respectively. UM- CE Upper Marker, LM- CE Lower Marker, VH- variable region PCR product, PP- unused Primer Pool, U- Unwanted products.