ChIP-qPCR Human IGX1A Negative Control

These Primers detect a specific sequence within a 1.4 Mb ORF-free intergenic region containing no known or predicted transcription start sites.
Genomic Position on Human Chromosome 12:
Chromosome RefSeq #	TSS Position	TSS Orientation	Assay Position
NC_000012.10	N/A	N/A	59208666
Note: These primers were designed from NCBI Homo sapiens Build Number: 37 Version: 1

Description

The ChIP-qPCR Negative Control Primers measure the amount of non-specific genomic DNA that co-precipitates during a ChIP procedure. The enclosed primers detect a specific genomic DNA sequence within an ORF-free intergenic region or "promoter desert" lacking any known or predicted structural genes. Both the amplification of a single product of the correct size and a high PCR efficiency are guaranteed when used with the qPCR master mixes. The uniform PCR efficiency and PCR conditions of all ChIP-qPCR Primers allow an accurate, scalable solution for multiple ChIP analyses. A threshold cycle value from specific ChIP fractions for these primers identical to any other primers with the non-immune serum ChIP fraction indicates similar levels of non-specific genomic DNA co-precipitation between ChIP fractions. Because each antibody interacts differently and to varying extents with non-specific DNA, a negative control should be performed for every experimental ChIP fraction. Include the ChIP-qPCR Negative Control Primers in your ChIP experiment to more accurately assess the specificity of your target binding enrichment.

Useful Links

Pricing and Ordering Technical Resources User Manual

Related ChIP-qPCR Products

qPCR Master Mixes Positive Control Primers Custom ChIP-qPCR Arrays

Brief Protocol: For Experienced Users

First time users should refer to the ChampionChIP PCR Primers User Manual.

Prepare ChIP DNA fractions. For each 25-μl PCR, mix the following components in a PCR tube: 12.5 μl RT² qPCR Master Mix, matched with your instrument 6.5 μl ddH2O 5.0 μl of either undiluted or diluted ChIP DNA template 1.0 μl ChIP-qPCR Primers (10 μM each) 25.0 μl final volume (Final primer concentration is 0.4 μM.) Place tubes in thermal cycler. Enter and run the following program: 95 ºC, 10 min1; 40 cycles of (95 ºC, 15 sec; 60 ºC, 60 sec2) 1The 10-minute step at 95 ºC is required to activate the HotStart DNA polymerase. 2Detect and record SYBR® Green fluorescence from every well at the end of the 60 ºC annealing / extension step of each cycle. Immediately follow the cycling program with your instrument's default melting curve program.

Materials Included / Packing List

Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt. One tube of ChIP-qPCR Primers (10 µM each primer, 2 primers, 200 µl) for 200 reactions One Certificate of Performance

Storage Conditions

The ChIP-qPCR Primers are shipped at ambient temperature. For long-term storage, store primers at -20 ºC.

Related Products

qPCR Master Mixes SYBR® is a registered trademark of Molecular Probes/Invitrogen.