Difference between revisions of "NMDA Receptors"
(→Primers for Davenport's Real-Time PCR experiment: added new primers designed in May 2009) |
|||
| Line 1: | Line 1: | ||
=Primers for Davenport's Real-Time PCR experiment= | =Primers for Davenport's Real-Time PCR experiment= | ||
| − | Primers were ordered 2009 March from Sigma Aldrich | + | Primers were ordered 2009 May from Sigma Aldrich |
| + | |||
| + | <tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top> | ||
| + | Probe Abbr Sequence Tm Size Genbank Reference | ||
| + | Rat NR1 subunit QNR1FOR ttcacagaagtgcgatctgg 60.0 108 U11418.1,2360-2467 PrimerPlus | ||
| + | QNR1REV atggacagggaaacgttctg 60.0 | ||
| + | |||
| + | Rat Serine Racemase QSERRACFOR tggtagatgcgttggtggta 60.0 103 NM_198757, 856-958 PrimerPlus | ||
| + | QSERRACREV tcagcagcgtacaccttcac 60.1 | ||
| + | |||
| + | Rat GADPH QGADPHFOR tcactgccactcagaagactg 60 ??? Andre designed with sequencher | ||
| + | QGADPHREV ggatgaccttgcccacagc 60 . . . | ||
| + | |||
| + | </tab> | ||
| + | |||
| + | Primers were ordered 2009 March from Sigma Aldrich (NB: these primers worked fine in a test RT-PCR, but Andre said the product sizes should be 100-150bp, and the Tm's should be closer together. So, we redesigned and ordered primers above). | ||
<tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top> | <tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top> | ||
| Line 13: | Line 28: | ||
Note: GADPHFOR and REV maybe mixed up . . . . | Note: GADPHFOR and REV maybe mixed up . . . . | ||
</tab> | </tab> | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
=Primers for Real-Time PCR= | =Primers for Real-Time PCR= | ||
Revision as of 15:06, 23 May 2009
Primers for Davenport's Real-Time PCR experiment
Primers were ordered 2009 May from Sigma Aldrich
| Probe Abbr Sequence Tm Size Genbank Reference |
|---|
| Rat NR1 subunit QNR1FOR ttcacagaagtgcgatctgg 60.0 108 U11418.1,2360-2467 PrimerPlus |
| QNR1REV atggacagggaaacgttctg 60.0 |
| Rat Serine Racemase QSERRACFOR tggtagatgcgttggtggta 60.0 103 NM_198757, 856-958 PrimerPlus |
| QSERRACREV tcagcagcgtacaccttcac 60.1 |
| Rat GADPH QGADPHFOR tcactgccactcagaagactg 60 ??? Andre designed with sequencher |
| QGADPHREV ggatgaccttgcccacagc 60 . . . |
Primers were ordered 2009 March from Sigma Aldrich (NB: these primers worked fine in a test RT-PCR, but Andre said the product sizes should be 100-150bp, and the Tm's should be closer together. So, we redesigned and ordered primers above).
| Probe Abbr Sequence Tm Size Genbank Reference |
|---|
| NR1 RTNR1FOR GTTCTTCCGCTCAGGCTTTG 66.0 66 RNU11418, 2396-2461 Yamamoto et al. 2008 PMID 17957211 |
| RTNR1REV AGGGAAACGTTCTGCTTCCA 65.8 |
| Serine Racemase RTSERRACFOR ATTGCAAGAAACTGGCCATC 63.9 295 NM198757, 664-958 Yoshikawa et al., 2006 PMID 16973158 |
| RTSERRACREV TCAGCAGCGTACACCTTCAC 64.1 |
| GADPH RTGADPHFOR TGCACCACCAACTGCTTAG 62.5 201 DQ403053.1, 397-597 Uno et al., 2002 PMID 12068078 |
| RTGADPHREV GGATGCAGGGATGATGTTC 63.0 |
| Note: GADPHFOR and REV maybe mixed up . . . . |
Primers for Real-Time PCR
Taken from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems
NR1
Forward primer 5'-GTTCTTCCGCTCAGGCTTTG-3' Reverse primer 5'-AGGGAAACGTTCTGCTTCCA-3' TaqMan probe 5'-FAM-CGGCATGCGCAAGGACAGCC-TAMRA-3'
NR2A
Forward primer 5'-AGCCCCCTTCGTCATCGTA-3' Reverse primer 5'-GACAGGGCACCGTGTTCCT-3' TaqMan probe 5'-FAM-AGGACATAGACCCCCTGACTGAGACCTGTG-TAMRA-3'
NR2B
Forward primer 5'-CCCCCAAGTTCTGGTTGGT-3' Reverse primer 5'-TTTTGGGAACGAGCTTTGCT-3' TaqMan probe 5'-FAM-TTGGCCGTCTTGGCCGTATCAGG-TAMRA-3'
NR2C
Forward primer 5'-GGGTGATGATGTTCGTGATGTG-3' Reverse primer 5'-GTGAGGTTCTGGTTGTAGCTGACA-3' TaqMan probe 5'-FAM-TCACGGTGGTTGCCATCACTGTCTTC-TAMRA-3'
Primers chosen for experiment (according to Andre's advice)
(designed using PrimerPlus3
NR1 U11418.1 2360 Forward ttcacagaagtgcgatctgg, Tm 60.0 2467 Reverse atggacagggaaacgttctg, Tm 60.0 Product size of 108 bp
Serine Racemase NM_198757
856 forward: tggtagatgcgttggtggta, Tm 60.0 958 reverse: tcagcagcgtacaccttcac, Tm60.1
674-807, product size 134bp
Rat GAPDH
Forward TCACTGCCACTCAGAAGACTG Reverse GGATGACCTTGCCCACAGC