|
|
| Transcripts Originating from
this RefSeq TSS:
|
| NCBI Gene Symbol
|
Gene ID |
RefSeq # |
| GAPDH
|
2597 |
NM_002046.3 |
| TSS Relative Positioning:
|
| Assay Tile
|
Assay Position |
| Proximal
|
(+)331 bp |
| Genomic Position on Human
Chromosome 12:
|
| Chromosome RefSeq #
|
TSS Position |
TSS Orientation |
Assay Position |
| NC_000012.10
|
6513918 |
Forward |
6514249 |
| NOTE: These primers were designed
from NCBI Homo sapiens Genome Build Number: 36 Version: 2
|
| Description |
|
The ChIP-qPCR Positive Control Primers insure the
effectiveness of your ChIP preparation procedure and provide a control for
successful chromatin immunoprecipitation. When used with a RNA Pol II ChIP DNA
fraction, the Primers are designed to measure the amount of basal transcriptional
machinery binding proximal to a constitutive promoter. The enclosed primers
detect genomic DNA within one kb of the transcription start site of a highly
expressed housekeeping gene where RNA Polymerase II binding should occur. 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. High levels of Pol II ChIP enrichment compared to a non-immune
ChIP enrichment, determined using the ChIP-qPCR Positive Control Primers, indicate
a successfully executed ChIP procedure. Include the ChIP-qPCR Positive Control in your ChIP experiment to insure the quality of
your ChIP DNA material and process.
|
| Related
ChIP-qPCR Products |
|
|
| 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.
|
|
