Difference between revisions of "NMDA Receptors"

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(added table of primers for RAD realtime experiment)
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=Primers for Davenport's Real-Time PCR experiment=
 
=Primers for Davenport's Real-Time PCR experiment=
  
Primers were ordered 2009 March from Sigma Aldrich
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Primers were ordered 2009 May from Sigma Aldrich
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 +
(designed using [https://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi PrimerPlus3])
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 +
 
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<tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top>
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Probe Abbr Sequence Tm Size Genbank Reference
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Rat NR1 subunit QNR1FOR ttcacagaagtgcgatctgg 60.0 108 U11418.1,2360-2467 PrimerPlus
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QNR1REV atggacagggaaacgttctg 60.0
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Rat Serine Racemase QSERRACFOR tggtagatgcgttggtggta 60.0 103 NM_198757, 856-958 PrimerPlus
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QSERRACREV tcagcagcgtacaccttcac 60.1
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Rat GADPH QGADPHFOR tcactgccactcagaagactg 60 123 DQ403053, 467-589 Andre designed with sequencher
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QGADPHREV ggatgaccttgcccacagc 60 . . .
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</tab>
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 +
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>
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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=
  
Taken from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems
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Looked at primers from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems, but ended up designing own primers above.
  
  
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'
 
  
 
[[Category:Taste_Aversion]]
 
[[Category:Taste_Aversion]]

Revision as of 09:05, 14 October 2021

Primers for Davenport's Real-Time PCR experiment

Primers were ordered 2009 May from Sigma Aldrich

(designed using PrimerPlus3)


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 60 123 DQ403053, 467-589 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

Looked at primers from Yamamoto et al. 2008 PMID 17957211 using Applied Biosystems, but ended up designing own primers above.