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Created page with "# Locate the potential gene regions you believe your protein is bound to (for glucocorticoid-induced ChIP-seq peaks refer to Locating ChIP-seq Peaks protocol). Make sure this..."
# Locate the potential gene regions you believe your protein is bound to (for glucocorticoid-induced ChIP-seq peaks refer to Locating ChIP-seq Peaks protocol). Make sure this the genetic sequence is species appropriate. If not, you can use the BLAT option from the UCSC website [https://genome.ucsc.edu/index.html] and choose the desired species from the drop-down menu.
# The genetic region entered for primer search should be around 400 bp.
# Go to NCBI primer design [http://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?ORGANISM=9606&INPUT_SEQUENCE=NM_001618.3]
# Enter your sequence in the first box.
# PCR product size should be set to 70-150bp.
# Make sure to select the proper species in the “organism” section.
# Click ‘Get Primers’.
# Choose a primer pair that is towards the middle region, if available.
# Like always you should print off this page and keep it in your records, indicating the primer pair you ordered, the NM# (if one is associated) and the sequences and locations of the forward and reverse primers. Remember to record these primers in the primer database as well.
# Order primers from IDT [https://www.idtdna.com/site]. It is important to indicate on the primer database and in the name on the label that these are ChIP primers so as not to be confused with normal qPCR primers.
# The genetic region entered for primer search should be around 400 bp.
# Go to NCBI primer design [http://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?ORGANISM=9606&INPUT_SEQUENCE=NM_001618.3]
# Enter your sequence in the first box.
# PCR product size should be set to 70-150bp.
# Make sure to select the proper species in the “organism” section.
# Click ‘Get Primers’.
# Choose a primer pair that is towards the middle region, if available.
# Like always you should print off this page and keep it in your records, indicating the primer pair you ordered, the NM# (if one is associated) and the sequences and locations of the forward and reverse primers. Remember to record these primers in the primer database as well.
# Order primers from IDT [https://www.idtdna.com/site]. It is important to indicate on the primer database and in the name on the label that these are ChIP primers so as not to be confused with normal qPCR primers.
