Sheet ID PS_05495
Release date : 5/9/2022
Print date : 5/9/2022

PCR protocol
RBRC 02096
PrimerStrain name B6.Cg-Tg(CAG-floxed Neo-EGFP)REP08Osb

primer_nameprimer_seq

lengthReference

1P3

GCAACGTGCTGGTTGTTGTGCTGTCTCATC

30Biochem. Biophys. Res. Commun. 321, 275–279 (2004).

2P4

GGACTTGAAGAAGTCGTGCTGC

22Biochem. Biophys. Res. Commun. 321, 275–279 (2004).

3mLC3ex3GT

TGAGCGAGCTCATCAAGATAATCAGGT

27Mol. Biol. Cell., 15, 1101-1111(2004).

4mLC3ex4AG

GTTAGCATTGAGCTGCAAGCGCCGTCT

27Mol. Biol. Cell., 15, 1101-1111(2004).

Reaction Components
Enzyme TaKaRa LA Taq with GC buffer (Cat. No. RR02AG)
Reagent
H2O
PCR bufferdNTP
primer 1
primer 2
primer 3
primer 4DNATaq

Reaction 1
2.9
10.0
2X
2.0
2.5 mM
1.5
10 pmol/μl
1.5
10 pmol/μl
0.5
10 pmol/μl
0.5
10 pmol/μl
1.0
0.1
5 U/μl2Conc

34

Reaction Conditions
temp
step (℃)
194
294（μL）
360
472
572

time
note(sec)
18030 *30 *180 **Repeat for 30 cycles
600

Marker 1 kb DNA ladder (Invitrogen, Cat. 15615-024)
Geltotalprimer_set

1% agarose in 1 X TAE (100 V, 30 min)20

product_size

product (allele type)primer 1

primer 2

2,100 bp

primer 3

primer 4

550 bpInternal control (LC3)primer 1

primer 2

300 bpTg (dNeo)

Comment Standard PCRTg

Sheet ID PS_01499
Release date : 3/1/2008
Print date : 5/9/2022

PCR protocol
RBRC 02096
PrimerStrain name B6.Cg-Tg(CAG-floxed Neo-EGFP)REP08Osb

primer_nameprimer_seq

lengthReference1

olMR0872

AAGTTCATCTGCACCACCG

19

JAX Genotyping Protocol

2#2

TGCTCAGGTAGTGGTTGTCG

20

Depositor’s primer

3

olMR0042

CTAGGCCACAGAATTGAAAGATCT24

Internal control (IL-2)

JAX Genotyping Protocol

4

olMR0043

GTAGGTGGAAATTCTAGCATCATCC

25

Internal control (IL-2)

JAX Genotyping Protocol

Reaction Components
Enzyme TaKaRa Ex Taq (Cat. No. RR001A)
Reagent
H2O
PCR bufferdNTP
primer 1
primer 2
primer 3
primer 4
DNATaqReaction 1
15.3
2.5
10 X
2.0
2.5 mM
1.0
10 pmol/μl
1.010 pmol/μl
1.010 pmol/μl
1.010 pmol/μl
1.00.22Conc

34Reaction Conditions
temp
step (℃)
194
294（μL）
360
472
572

time
note(sec)180
30 *30 *60 **Repeat for 30 cycles
300

Marker 100 bp ladder (Invitrogen, Cat. 15628-050)
Geltotalprimer_set

25

product_size

primer 1

primer 2475 bp

primer 3

primer 4324 bp

Comment Standard PCR

2% agarose in 1 X TAE (100 V, 30 min)product (allele type)
Tg
Internal control (IL-2)

Sheet ID PS_01000
Release date : 10/2/2007
Print date : 5/9/2022

PCR protocol
RBRC 02096
PrimerStrain name B6.Cg-Tg(CAG-floxed Neo-EGFP)REP08Osb

primer_nameprimer_seq

lengthReference

1A

GAGCTAGCCACCATGGTGAGCAAGGGCGAG

30

Depositor’s primer

2BTCACCTTGATGCCGTTCTTCT

21

Depositor’s primer

3

olMR0042

CTAGGCCACAGAATTGAAAGATCT

24

Internal control (IL-2)

JAX Genotyping Protocol

4

olMR0043

GTAGGTGGAAATTCTAGCATCATCC

25

Internal control (IL-2)

JAX Genotyping Protocol

Reaction Components
Enzyme TaKaRa Ex Taq (Cat. No. RR001A)
Reagent
H2O
PCR bufferdNTP
primer 1
primer 2
primer 3
primer 4
DNATaqReaction 1
15.32.510 X
2.0
2.5 mM
1.0
10 pmol/μl
1.0
10 pmol/μl
1.0
10 pmol/μl
1.0
10 pmol/μl
1.00.2

2Conc

34Reaction Conditions
temp
step (℃)
194
294（μL）
360
472
572

time
note(sec)180
30 *30 *60 **Repeat for 30 cycles
300

Marker 100 bp ladder (Invitrogen, Cat. 15628-050)
Gel
total

primer_set

25

product_size

primer 1

primer 2500 bp

primer 3

primer 4

324 bpComment Standard PCR

2% agarose in 1 X TAE (100 V, 30 min)product (allele type)
Tg
Internal control (IL-2)