ERRATUM Jacobson, D.R. and T. Moskovits. 1991. Rapid, nonradioactive screening for activating ras oncogene mutations using PCR-primer introduced restriction analysis (PCR-PIRA). PCR Methods Applic. l : 146-148.
TABLE 1A Screening PCR Primers and Restriction Sites Formed Codon
Name
DNA
Primers and Sites Created
i0 12
KS12
Restriction enzyme
Sequence ii 12
I \/ K-Ras
The footnote to Table 1 was inadvertently omitted. The correct version of the table is reproduced here.
Screening Restriction
\I \
5'AACTTGTGGTAGTTGGAG~_~t... i0 ii
I \/ N-Ras
12
NSI2
\I
\
13
KS13
5 'T T G T G G T A G T T C C A G C T G G T ~ g c . . .
N-Ras
13
NSI3
5'TGGTGGTGGTTCCAGCAGGT~yt...
ii 12
K-Ras (CA-)
61 KS61
I
13
K-Ras
I \/
Msp
\ gt...
12
I \/
I
12
\I
5 'A A C T G G T G G T G G T T G G A G ~ _ Ii
Msp
Pflm
I
Pflm
I
13
\/
\
59
60 61
I \I
\/
\
5'ATATTCTCGACACAGCTC.ATcaa...
Bcl
I
Msc
I
Mse
I
Mse
I
59 60 61 N-Ras (CA-)
61
K-Ras
61
I \/ NS61
(--A)
KS61S
\/
59
60 61
/ \/
\/
\
5'ATTCTCGACACAGCAGGTT_Aa... 59 60
N-Ras (--A)
61
I \I \ /
61
9 NS61S
\
5'ACATAOTGGATACAG~Ccaa...
\
5 ! ATACTGGATACAGCTGGTTA&...
TABLE 1B PCR Primers Used to Determine W h e t h e r Mutations at the Codon 61 Position 3 Are Activating ( A ~ C or A ~ T ) or Silent ( A ~ G ) Codon
Screening Restriction Name
DNA
Primers and Sites Created
Sequence 63
/ \/ K-Ras (--S)
N-Ras (--G)
61
61
KS61G
NS61G
Restriction enzyme
62
\/
61
\
5 'C T C A T T G C A C T G T A C A G C T C C t g . . . 63
62 61
I \I
\I
Alwn
I
Alwn
I
\
5'CTCATGGCACTGTACAGTTCCtg...
Primer sequences are capitalized; lowercase letters indicate bases incorporated during PCR. Bases m i s m a t c h e d from the genomic DNA to create new restriction sites are underlined. Bases comprising the new sites are in boldface. In Table 1B the base being screened is marked with an asterisk.
1:2999
by Cold Spring Harbor Laboratory Press ISSN 1054-9803/92 $3.00
IJ~R Methods and Applications 299
TABLE 1A Screening PCR Primers and Restriction Sites Formed Codon
Name
DNA
Primers and Sites Created
i0 12
KS12
Restriction enzyme
Sequence ii 12
I \/ K-Ras
The footnote to Table 1 was inadvertently omitted. The correct version of the table is reproduced here.
Screening Restriction
\I \
5'AACTTGTGGTAGTTGGAG~_~t... i0 ii
I \/ N-Ras
12
NSI2
\I
\
13
KS13
5 'T T G T G G T A G T T C C A G C T G G T ~ g c . . .
N-Ras
13
NSI3
5'TGGTGGTGGTTCCAGCAGGT~yt...
ii 12
K-Ras (CA-)
61 KS61
I
13
K-Ras
I \/
Msp
\ gt...
12
I \/
I
12
\I
5 'A A C T G G T G G T G G T T G G A G ~ _ Ii
Msp
Pflm
I
Pflm
I
13
\/
\
59
60 61
I \I
\/
\
5'ATATTCTCGACACAGCTC.ATcaa...
Bcl
I
Msc
I
Mse
I
Mse
I
59 60 61 N-Ras (CA-)
61
K-Ras
61
I \/ NS61
(--A)
KS61S
\/
59
60 61
/ \/
\/
\
5'ATTCTCGACACAGCAGGTT_Aa... 59 60
N-Ras (--A)
61
I \I \ /
61
9 NS61S
\
5'ACATAOTGGATACAG~Ccaa...
\
5 ! ATACTGGATACAGCTGGTTA&...
TABLE 1B PCR Primers Used to Determine W h e t h e r Mutations at the Codon 61 Position 3 Are Activating ( A ~ C or A ~ T ) or Silent ( A ~ G ) Codon
Screening Restriction Name
DNA
Primers and Sites Created
Sequence 63
/ \/ K-Ras (--S)
N-Ras (--G)
61
61
KS61G
NS61G
Restriction enzyme
62
\/
61
\
5 'C T C A T T G C A C T G T A C A G C T C C t g . . . 63
62 61
I \I
\I
Alwn
I
Alwn
I
\
5'CTCATGGCACTGTACAGTTCCtg...
Primer sequences are capitalized; lowercase letters indicate bases incorporated during PCR. Bases m i s m a t c h e d from the genomic DNA to create new restriction sites are underlined. Bases comprising the new sites are in boldface. In Table 1B the base being screened is marked with an asterisk.
1:2999
by Cold Spring Harbor Laboratory Press ISSN 1054-9803/92 $3.00
IJ~R Methods and Applications 299
