. Author manuscript; available in PMC: 2009 May 5.

Published in final edited form as: J Immunol Methods. 2006 Sep 5;316(1-2):59–66. doi: 10.1016/j.jim.2006.08.002

Table 1.

SHM in V_36-65 region in hybridomas^a

	Control	AID transfected (A)

	L27	L25A1	L27A8
Mutated sequences/total sequences (%)	1/24 (4.2%)	12/26 (46.1%)	12/23 (52.2%)
Mutation frequency^b	1.1×10⁻⁴	1.7×10⁻³	1.6×10⁻³
G:C mutations/total mutations	1/1 (100%)	16/16 (100%)	14/14 (100%)
Ts^c/total mutations	1/1 (100%)	13/16 (81.2%)	10/14 (71.4%)
Hs^d/total mutations	1/1 (100%)	13/16 (81.2%)	9/14 (64.3%)

Hybridomas transfected with AID were grown for 2 months. Heavy-chain V regions were amplified with Pfu polymerase, cloned in pCR blunt TOPO and sequenced. The region sequenced corresponds to 36-65 and is 363 bp long. The consensus sequence is available from Genbank accession no. M19292. Duplicated mutations were scored only once. V regions that shared the same mutation were scored as only one mutated sequence.

Frequency corresponds to mutations per base pair sequenced.

Ts: transition mutations

Hs: hotspots (mutations at G or C nucleotides within RGYW and WRCY).