Primers available for Premium Service

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Primer Name	Primer Express Tm @ 50nM, 50mM NaCl	Primer Sequence 5'-3'	size (nt)	Primer Dimer? (by Primer Express)
BGH_Reverse_Long	64.1 ºC	ACTAGAAGGCACAGTCGAGGCTGATC	26	Possible
BGH_Reverse_Core	61.2 ºC	TAGAAGGCACAGTCGAGGCTGAT	23	Not likely
BGH_Reverse_Invitrogen	50.6 ºC	TAGAAGGCACAGTCGAGG	18	Not likely
M13/pUC_Forward	53.7 ºC	TGTAAAACGACGGCCAGT	18	Yes
M13/pUC_Forward_23	69.8 ºC	CGCCAGGGTTTTCCCAGTCACGA	23	Not likely
M13/pUC_Reverse	54.1 ºC	TCACACAGGAAACAGCTATGAC	22	Not likely
M13/pUC_Reverse_GC+	54.3 ºC	GCTATGACCATGATTACGCC	20	Not likely
SP6_Promega	35.5 ºC	TATTTAGGTGACACTATAG	19	Not likely
SP6_NEB_24	47.0 ºC	CATACGATTTAGGTGACACTATAG	24	Not likely
SP6_Core	43.5 ºC	ATTTAGGTGACACTATAGAATAC	23	Not likely
T7 Promoter (Promega)	43.4 ºC	TAATACGACTCACTATAGGG	20	Possible
T7 Promoter Neo (Core)	38.4 ºC	TAATACGACTCACTATAGG	19	Not likely
T7_Promoter_EEV_Promega		AAGGCTAGAGTACTTAATACGA	22
T7_Terminator	52.6 ºC	GCTAGTTATTGCTCAGCGG	19	Possible
T3_Promega	49.4 ºC	ATTAACCCTCACTAAAGGGA	20	Not likely

Primer Comments

• BGH primers anneal to pKPPURO, pNKTV1907, pcDNA3, pcDNA3.1, pcDNA3.1/Neo, pCR3.1, pIRE51hyg, pIRE51 neo
• M13R/pUC Reverse (=M13R) primer does have some homology with common vectors at the 3' end and can produce some background.
  • M13/pUC_Reverse_GC+ (M13R GC+) primer was designed to avoid this problem and works with most pUC-, pGEM-, and pBluescript-derived plasmids. However, there are some vectors on which it will not work due to different sequence at 3’ and / or 5’ ends
  • M13 R primer anneals to some plasmids at the primers 3’ end 16 bp or greater, but not the full length.
• SP6 primer does not match with sequence at the 3’ end of the SP6 genome perfectly. Therefore, the different SP6 primers do not match all vectors with "SP6" primer sites as the sequences differ slightly.
• T7 promoter Promega primer does not match 8 out of many other plasmids at the 3’ end.
  • T7 promoter Neo (core) primer can be used to sequence both the vectors with normal T7 promoter sequence and pCIneo-derived vectors with the shorter T7 sequence which have two Gs at 3' end; whereas T7 primers has three Gs.
  • T7 promoter EEV Promega is utilized by Promega to sequence pCIneo derived vectors and the inserts they carry.

Vector Comments

• Invitrogen pCRII vector (had SP6 and T7 primer sites). Due to a Promega patent on the SP6 sequence, Invitrogen had to remove the SP6 site to make the pCR2.1 vector, which does not have an SP6 site.

Before specifying a particular primer for DNA sequencing, verify the primer site sequence is present in your vector.