I am using gefitinib to inhibit the EGFR. I want to perform RT qPCR on EGFR target genes to make sure the drug is actually working.
Hi Cora,
I have never done RT qPCR on EGFR or its targets, but perhaps one of the following two websites can help you create your own primer pairs:
https://primerdepot.nci.nih.gov/
http://www.ncbi.nlm.nih.gov/tools/primer-blast/
Good luck
Your MEKK1 sequence is:
>gi|2815887|gb|AF042838.1| Homo sapiens MEK kinase 1 (MEKK1) mRNA, partial cds
CCGAGCCCTGAGGCAGGCGGCGGCGGAGGAGCCCTCAAGGCGAGCAGCGCGCGCGCGGCTGCCGCGGGAC
TGCTGCGGGAGGCGGGCAGCGGGGGCCGCGAGCGGGCGGACTGGCGGCGGCGGCAGCTGCGCAAAGTGCG
GAGTGTGGAGCTGGACCAGCTGCCTGAGCAGCCGCTCTTCCTTGCCGCCTCACCGCCGGCCTCCTCGACT
TCCCCGTCGCCGGAGCCCGCGGACGCAGCGGGGAGTGGGACCGGCTTCCAGCCTGTGGCGGTGCCGCCGC
CCCACGGAGCCGCCAGCCGGCGCGGCGCCCACCTTACCGAGTCGGTGGCGGCGCCGGACAGCGGCGCCTC
GAGTCCCGCAGCGGCCGAGCCCGGGGAGAAGCGGGCGCCCGCCGCCGAGCCGTCTCCTGCAGCGGCCCCC
GCCGGTCGTGAGATGGAGAATAAAGAAACTCTCAAAGGGTTGCACAAGATGGATGATCGTCCAGAGGAAC
GAATGATCAGGGAGAAACTGAAGGCAACCTGTATGCCAGCCTGGAAGCACGAATGGTTGGAAAGGAGAAA
TAGGCGAGGGCCTGTGGTGGTAAAACCAATCCCAGTTAAAGGAGATGGATCTGAAATGAATCACTTAGCA
GCTGAGTCTCCAGGAGAGGTCCAGGCAAGTGCGGCTTCACCAGCTTCCAAAGGCCGACGCAGTCCTTCTC
CTGGCAACTCCCCATCAGGTCGCACAGTGAAATCAGAATCTCCAGGAGTAAGGAGAAAAAGAGTTTCCCC
AGTGCCTTTTCAGAGTGGCAGAATCACACCACCCCGAAGAGCCCCTTCACCAGATGGCTTCTCACCATAT
AGCCCTGAGGAAACAAACCGCCGTGTTAACAAAGTGATGCGGGCCAGACTGTACTTACTGCAGCAGATAG
GGCCTAACTCTTTCCTGATTGGAGGAGACAGCCCAGACAATAAATACCGGGTGTTTATTGGGCCTCAGAA
CTGCAGCTGTGCACATGGAACATTCTGTATTCATCTGCTATTTGTGATGCTCCGGGTGTTTCAACTAGAA
CCTTCAGACCCAATGTTATGGAGAAAAACTTTAAAGAATTTTGAGGTTGAGAGTTTGTTCCAGAAATATC
ACAGTAGGCGTAGCTCAAGGATCAAAGCTCCATCTCGTAACACCATCCAGAAGTTTGTTTCACGCATGTC
AAATTCTCATACATTGTCATCATCTAGTACTTCTACATCTAGTTCAGAAAACAGCATAAAGGATGAAGAG
GAACAGATGTGTCCTATTTGCTTGTTGGGCATGCTTGATGAAGAAAGTCTTACAGTGTGTGAAGACGGCT
GCAGGAACAAGCTGCACCACCACTGCATGTCAATTTGGGCAGAAGAGTGTAGAAGAAATAGAGAACCTTT
AATATGTCCCCTTTGTAGATCTAAGTGGAGATCTCATGATTTCTACAGCCACGAGTTGTCAAGTCCTGTG
GATTCCCCTTCTTCCCTCAGAGCTGCACAGCAGCAAACCGTACAGCAGCAGCCTTTGGCTGGATCACGAA
GGAATCAAGAGAGCAATTTTAACCTTACTCATTATGGAACTCAGCAAATCCCTCCTGCTTACAAAGATTT
AGCTGAGCCATGGATTCAGGTGTTTGGAATGGAACTCGTTGGCTGCTTATTTTCTAGAAACTGGAATGTG
AGAGAGATGGCCCTCAGGCGTCTTTCCCATGATGTCAGTGGGGCCCTGCTGTTGGCAAATGGGGAGAGCA
CTGGAAATTCTGGGGGCAGCAGTGGAAGCAGCCCGAGTGGGGGAGCCACCAGTGGGTCTTCCCAGACCAG
TATCTCAGGAGATGTGGTGGAGGCATGCTGCAGCGTTCTGTCAATGGTCTGTGCTGACCCTGTCTACAAA
GTGTACGTTGCTGCTTTAAAAACATTGAGAGCCATGCTGGTATATACTCCTTGCCACAGTTTAGCGGAAA
GAATCAAACTTCAGAGACTTCTCCAGCCAGTTGTAGACACCATCCTAGTCAAATGTGCAGATGCCAATAG
CCGCACAAGTCAGCTGTCCATATCAACACTGTTGGAACTGTGCAAAGGCCAAGCAGGAGAGTTGGCAGTT
GGCAGAGAAATACTAAAAGCTGGATCCATTGGTATTGGTGGTGTTGATTATGTCTTAAATTGTATTCTTG
GAAACCAAACTGAATCAAACAATTGGCAAGAACTTCTTGGCCGCCTTTGTCTTATAGATAGACTGTTGTT
GGAATTTCCTGCTGAATTTTATCCTCATATTGTCAGTACTGATGTTTCACAAGCTGAGCCTGTTGAAATC
AGGTATAAGAAGCTGCTGTCCCTCTTAACCTTTGCTTTGCAGTCCATTGATAATTCCCACTCAATGGTTG
GCAAACTTTCCAGAAGGATCTACTTGAGTTCTGCAAGAATGGTTACTACAGTACCCCATGTGTTTTCAAA
ACTGTTAGAAATGCTGAGTGTTTCCAGTGTTTCCACTCACTTCACCAGGATGCGTCGCCGTTTGATGGCT
TATGCAGATGAGGTGGAAATTGCCGAAGCCATCCAGTTGGGCGTAGAAGACACTTTACAACGACAACAAC
ACAACAGCTTTTGCAGGCATCTGTTCCCAACAACTATCTGGAAACCACAGAGAACAGTTCCCCTTGAGTG
CACAGTCCATTTAGAGAAAACTGGAAAAGGATTATGTGCTACAAAATTGAGTGCCAGTTCAGAGGACATT
TCTGAGAGACTGGCCAGGATTTCAGTAGGACCTTCTAGTTCAACAACAACAACAACAACAACAACAGAGC
AACCAAAGCCAATGGTTCAAACAAAAGGCAGACCCCACAGTCAGTGTTTGAACTCCTCTCCTTTATCTCA
TCATTCCCAATTAATGTTTCCAGCCTTGTCAACCCCTTCTTCTTCTACCCCATCTGTACCAGCTGGCACT
GCAACAGATGTCTCTAAGCATAGACTTCAGGGATTCATTCCCTGCAGAATACCTTCTGCATCTCCTCAAA
CACAGCGCAAGTTTTCTCTACAATTCCACAGAAACTGTCCTGAAAACAAAGACTCAGATAAACTTTCCCC
AGTCTTTACTCAGTCAAGACCCTTGCCCTCCAGTAACATACACAGGCCAAAGCCATCTCGACCTACCCCA
GGTAATACAAGTAAACAGGGAGATCCCTCAAAAAATAGCATGACACTTGATCTGAACAGTAGTTCCAAAT
GTGATGACAGCTTTGGCTTGAGCAGCAATAGTAGTAATTGCTGTTATACCAGTGACGAGACAGTGTTCAC
CCCAGTAGAGGAGAAATGCAGATTAGATGTCAATACAGAGCTCAACTCCAGTATTGAGGACCTTCTTGAA
GCATCTATGCCTTCAAGTGATACAACAGTAACTTTTAAGTCAGAAGTTGCTGTCCTGTCTCCTGAAAAGG
CTGAAAATGATGATACCTACAAAGATGATGTGAATCATAATCAAAAGTGCAAAGAGAAGATGGAAGCTGA
AGAAGAAGAAGCTTTAGCAATTGCCATGGCAATGTCAGCGTCTCAGGTAGCCCTCCCCATAGTTCCTCAG
CTGCAGGTTGAAAATGGAGAAGATATCATCATTATTCAACAGGATACACCAGAGACTCTACCAGGACATA
CCAAAGCAAAACAACCGTATAGAGAAGACACTGAATGGCTGAAAGGTCAACAGATAGGCCTTGGAGCATT
TTCTTCTTGTTATCAGGCTCAAGATGTGGGAACTGGAACTTTAATGGCTGTTAAACAGGTGACTTATGTC
AGAAACACATCTTCTGAGCAAGAAGAAGTAGTAGAAGCACTAAGAGAAGAGATAAGAATGATGAGCCATC
TGAATCATCCAAACATCATTAGGATGTTGGGAGCCACGTGTGAGAAGAGCAATTACAATCTCTTCATTGA
ATGGATGGCAGGGGGATCGGTGGCTCATTTGCTGAGTAAATATGGAGCCTTCAAAGAATCAGTAGTTATT
AACTACACTGAACAGTTACTCCGTGGACTTTCGTATCTCCATGAAAACCAAATCATTCACAGAGATGTCA
AAGGTGCCAATTTGCTAATTGACAGCACTGGTCAGAGACTAAGAATTGCAGATTTTGGAGCTGCAGCCAG
GTTGGCATCAAAAGGAACTGGTGCAGGAGAGTTTCAGGGACAATTACTGGGGACAATTGCATTTATGGCA
CCTGAGGTACTAAGAGGTCAACAGTATGGAAGGAGCTGTGATGTATGGAGTGTTGGCTGTGCTATTATAG
AAATGGCTTGTGCAAAACCACCATGGAATGCAGAAAAACACTCCAATCATCTTGCTTTGATATTTAAGAT
TGCTAGTGCAACTACTGCTCCATCGATCCCTTCACATTTGTCTCCTGGTTTACGAGATGTGGCTCTTCGT
TGTTTAGAACTTCAACCTCAGGACAGACCTCCATCAAGAGAGCTACTGAAGCATCCAGTCTTTCGTACTA
CATGGTAGCCAATTATACAGATCAACTACGTAGAAACAGGATGCTCAACAAGAGAAAAAAAACTTGTGGG
GAACCACATTGATATCTACGGCCATGATGCCACTGAACAGCTATGAACGAGGCCAGTGGGGAACCCTTAC
CTAAGTATGTGATTGACAAATCATGATCTGTACCTAAGCTCAGTATGCAAAAGCCCAAACTAGTGCAGAA
ACT
Select the portion of the sequence you want to reproduce and apply the primer program: http://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?ORGANISM=9606&INPUT_SEQUENCE=AF042838.1&LINK_LOC=nuccore
This procedure will give you the adequate primers
07 August 2017 3,203 1 View
Because of logistics I have to store my stained and fixed cells for 3 days before flow cytometry. Has this ever worked for anyone? I am used to staining and analysing on the same day...
04 May 2016 4,047 4 View
Hi. Please tell me what guidelines should i need to follow for questionaries' type research work in India. It is not hospital based work, we are conduction basic institutional based qualitative...
03 March 2021 2,037 3 View
Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...
03 March 2021 5,999 3 View
Hello all, I'm currently working on my undergraduate thesis and I'm having difficulties trying to access KLD Stats and Sustainalytics because my campus Airlangga University and apparently other...
02 March 2021 8,525 1 View
Hi everyone, I'm studying Marketing and I would like to write my PhD thesis on the topic of pricing. Any specific ideas?
02 March 2021 9,706 5 View
02 March 2021 3,060 3 View
02 March 2021 932 1 View
how to calculate BEHAVIOURAL REGULATION IN EXERCISE QUESTIONNAIRE BREQ 3 ,which contain 24 questions and each question has 5 answers (0- Strongly disagree , 1-disagree, 2- Neither agree nor...
02 March 2021 811 3 View
I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...
02 March 2021 9,945 3 View
Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...
02 March 2021 3,024 2 View
Please provide Book Title and author name
02 March 2021 9,059 3 View