|
TCRBV amplifications1
|
IGHV amplifications2
|
|---|
|
No.:
|
Primer:
|
Sequence:
|
No.:
|
Primer:
|
Sequence
|
|---|
|
1
|
VβI
|
tccatgctcttctgctgtgt
|
10
|
FR1-5
|
gaggagaagctggtggagt
|
|
2
|
VβIV
|
kgcaycgggstkctctg
|
11
|
FR1-3
|
ctcctgtgtcggctctgga
|
|
3
|
VβVII
|
ctcascggramcctttgc
|
12
|
FR3
|
tgagaaccgaagacacggc
|
|
4
|
FR3βIa
|
gactchgchstgtwyytctgtg
|
13
|
α-Cμ
|
gggacgaagatgttcaagac
|
|
5
|
FR3βIb
|
crgacatctvkrtayytstgtg
|
14
|
α-Cδ
|
gctgggagctgccgagat
|
|
6
|
FR3βIV
|
gactcsgcygtgtatctctg
|
15
|
α-Cγ
|
ccgtccacgtaccaggagaa
|
|
7
|
FR3βVII
|
gactcrgccacctacstctg
|
16
|
α-Cα
|
gagcccaggagcaggtct
|
|
8
|
α-Cβ1
|
tctccgcttccgatggttca
|
17
|
α-Cε
|
gtccggatggtggtgtttg
|
|
9
|
α-Cβ2
|
gtggtctcacctgctgcag
|
18
|
α-JH
|
tgaggacacgacgacttcaa
|
- 1Following primer pairs (stated in brackets for 1st, 2nd and 3rd PCR respectively) were used for TCRBV amplifications: VβI-VβIII families (1/8, 1/9, 4 + 5/9), VβIV-VβVI families (2/8, 2/9, 6/9) and VβVII family (3/8, 3/9, 7/9). Annealing temperatures (Tm) were 54 °C for 1st PCR, 61 °C for 2nd PCR and 55 °C for 3rd PCR.
- 2Following primer pairs (stated in brackets for 1st, 2nd and 3rd PCR respectively) were used for IGHV amplifications: total immunoglobulins (10/18, 11/18, 12/18), IgM (10/13, 11/13, 12/18), IgD (10/14, 11/14, 12/18), IgG (10/15, 11/15, 12/18), IgA (10/16, 11/16, 12/18) and IgE (10/17, 11/17, 12/18). Annealing temperatures (Tm) were 58 °C for 1st and 2nd PCR and 55 °C for 3rd PCR.
