Sybr green dynamo hs kit

Total RNA was extracted from 5 × 10⁶ B cells using the RNeasy Mini Kit (Qiagen). First-strand cDNA was synthesized from 2 μg RNAs using the SuperScript^™ III System (Invitrogen) and analyzed by qPCR using SYBR Green (Dynamo HS kit; New England Biolabs) and appropriate primers (Table 2). PCR was performed in a MyiQ Real-Time PCR System (Bio-Rad Laboratories) according to the following protocol: 95°C for 30 s, 40 cycles of 95°C for 10 s, 60°C for 30 s, 72°C for 30 s. Melting curve analysis was performed at 72°C–95°C. The ΔΔCt method was used to analyze levels of transcripts and data were normalized to the level of Cd79b, which encodes the BCR Igβ chain constitutively expressed in B cells.

Total RNA was extracted from 5 × 10⁶ B cells using the RNeasy Mini Kit (Qiagen). First-strand cDNA was synthesized from 2 μg RNAs using the SuperScript^™ III System (Invitrogen) and analyzed by qPCR using SYBR Green (Dynamo HS kit; New England Biolabs) and appropriate primers (Supplemental Table S2). PCR was performed in a MyiQ Real-Time PCR System (Bio-Rad Laboratories) according to the following protocol: 95°C for 30 s, 40 cycles of 95°C for 10 s, 60°C for 30 s, 72°C for 30 s. Melting curve analysis was performed at 72°C–95°C. The ΔΔCt method was used to analyze levels of transcripts and data were normalized to the level of Cd79b, which encodes the BCR Igβ chain constitutively expressed in B cells.