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PerfectHyb� Hybridization Solution

This is a premixed hybridization solution for Northern and Southern Analyses containing a hybridization enhancer. It provides simpler, more rapid analysis than with conventional methods. Precipitation occurs easily in some commercially available hybridization solutions due to high viscosity or the addition of salmon sperm DNA. However, PefectHyb� does not require additives and shows no precipitation occurrence at normal temperatures due to low viscosity, making handling simple. Moreover, it is suitable for analysis of non-RI as well as RI-labeled probes such as DIG.

Application:

Northern and Southern Blot Analysis
This solution requires no additives so handling is easy.

1. Northern Blot Analysis Using PefectHyb
Northern blot analysis was performed with 5 �g of total RNA from HeLa cells. Total RNA was extracted using MagExtractor-RNA- (Code No. NPK-201), and then electrophoresis performed in acrylamide gel containing 50% formamide for transfer to the membrane.
An internal probe was prepared from [32P]dCTP with transferrin receptor gene as the template using a Prime-ItII Random Primer Labeling Kit (Code No. SC300385). It was then purified with MagExtractor-PCR & Gel Clean up- (Code No. NPK-601).
Detection was performed with PefectHyb and other commercially available hybridization solutions and by the conventional method to compare results. The results showed that PefectHyb provided higher sensitive detection than other solutions

2. Southern Blot Using PefectHyb
Human genomic DNA VNTR was analyzed.
1 �g Genomic DNA extracted from a human cell line and digested with Hae? was electrophoresed and transferred to a membrane. Dig-labeled polymorphic analysis probe YNH24 (D2S44) was used for detection.PefectHyb was confirmed to have higher sensitivity and specificity than other companies� solutions.

Feature and Advantages:

  • High Sensitivity
    This solution contains a hybridization enhancer and generates a strong signal, making it effective for the detection of genes with low expression rates.
  • Optimize Performance of a Broad Range of Applications
    This solution can be used in Northern and Southern blot analysis. It is also ideal for non-RI analysis.
  • Rapid
    Using this solution allows completion of normal hybridization in 1-2 hours.
  • Easy to use
    This solution does not develop crystal precipitation and has a low viscosity at normal temperatures, allowing simple handling. Addition of salmon sperm DNA is also not necessary. Moreover, it allows hybridization and washing procedures at the same temperature and is thus ideal for analysis using hybridization ovens.

One-Point Advice:

  • Use for the mRNA Blotting Membrane
    We recommend that RI-labeled cDNA probes or oligo probes be used in analysis of pre-blotting membranes in consideration of background etc.
  • Detection of the Gene Navigator cDNA Array System cDNA array
    The use of PefectHyb� Hybridization Solution -filter array- is recommended for detection of the above-mentioned product.

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PerfectHyb Hybridization Solution TYB-HYB-101 250 ML �355

 

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