A multiplexed method of detecting an analyte in a sample using two or more padlock probes each specific to a different target sequence, the target sequence being either part of an analyte or indicative of the presence of an analyte in the sample, wherein each padlock probe comprises: (i) 5' and 3' end sequences capable of hybridising to a different target nucleic acid sequence such that upon said hybridisation the ends of the padlock probe are brought into juxtaposition, directly or indirectly, for ligation to circularise the padlock probe; and, between the said 5' and 3' end sequences: (ii) at least one reporter sequence, at least one of which is an analyte-specific reporter sequence; (iii) a restriction oligonucleotide sequence capable of hybridising to a complementary sequence to create a restriction cleavage site, wherein any RO sequence in the probe is located 3' of the analyte-specific reporter sequence; said method comprising: (a) contacting the sample with the two or more padlock probes and allowing the probes to hybridise to their respective target sequences, if present; (b) circularising any padlock probe which has hybridised to its target nucleic acid sequence by ligation to form a rolling circle amplification template; (c) amplifying ligated padlock probes by performing at least a linear RCA reaction using the RCA template(s) formed in step (b) to form first rolling circle amplification product(s), wherein a first RCP is a concatemer comprising monomers which are complementary to the circularised padlock probe which templated its formation, and optionally, simultaneously with or subsequently to said linear RCA reaction, performing a further amplification reaction to amplify the first RCP(s); (d) performing, during or after step (c), a restriction cleavage step using a restriction enzyme capable of cleaving the restriction cleavage site, optionally together with a restriction oligonucleotide which is complementary to and capable of hybridising to the restriction oligonucleotide sequence, wherein said cleavage occurs 3' of the analyte-specific reporter sequence; and (e) detecting the amplification product(s) to detect the analyte(s),. (i) 5' and 3' end sequences capable of hybridising to a different target nucleic acid sequence such that upon said hybridisation the ends of the padlock probe are brought into juxtaposition, directly or indirectly, for ligation to circularise the padlock probe; and, between the said 5' and 3' end sequences: (ii) at least one reporter sequence, at least one of which is an analyte-specific reporter sequence; (iii) a restriction oligonucleotide sequence capable of hybridising to a complementary sequence to create a restriction cleavage site, wherein any RO sequence in the probe is located 3' of the analyte-specific reporter sequence; said method comprising: (a) contacting the sample with the two or more padlock probes and allowing the probes to hybridise to their respective target sequences, if present; (b) circularising any padlock probe which has hybridised to its target nucleic acid sequence by ligation to form a rolling circle amplification template; (c) amplifying ligated padlock probes by performing at least a linear RCA reaction using the RCA template(s) formed in step (b) to form first rolling circle amplification product(s), wherein a first RCP is a concatemer comprising monomers which are complementary to the circularised padlock probe which templated its formation, and optionally, simultaneously with or subsequently to said linear RCA reaction, performing a further amplification reaction to amplify the first RCP(s); (d) performing, during or after step (c), a restriction cleavage step using a restriction enzyme capable of cleaving the restriction cleavage site, optionally together with a restriction oligonucleotide which is complementary to and capable of hybridising to the restriction oligonucleotide sequence, wherein said cleavage occurs 3' of the analyte-specific reporter sequence; and (e) detecting the amplification product(s) to detect the analyte(s),. (a) contacting the sample with the two or more padlock probes and allowing the probes to hybridise to their respective target sequences, if present; (b) circularising any padlock probe which has hybridised to its target nucleic acid sequence by ligation to form a rolling circle amplification template; (c) amplifying ligated padlock probes by performing at least a linear RCA reaction using the RCA template(s) formed in step (b) to form first rolling circle amplification product(s), wherein a first RCP is a concatemer comprising monomers which are complementary to the circularised padlock probe which templated its formation, and optionally, simultaneously with or subsequently to said linear RCA reaction, performing a further amplification reaction to amplify the first RCP(s); (d) performing, during or after step (c), a restriction cleavage step using a restriction enzyme capable of cleaving the restriction cleavage site, optionally together with a restriction oligonucleotide which is complementary to and capable of hybridising to the restriction oligonucleotide sequence, wherein said cleavage occurs 3' of the analyte-specific reporter sequence; and (e) detecting the amplification product(s) to detect the analyte(s),.