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YellowSub

Yellow SubTM

Yellow SubTM, the Novel PCR Additive for Enhanced Performance

Yellow SubTM is suitable for many enzymatic reactions. It increases the yield and specificity of your PCR® - and saves time. Just add the specially formulated 10 x Yellow SubTM to your PCR®-mix. Yellow SubTM also contains a loading buffer and dye, so after the reaction you can apply the PCR®-mix directly on your agarose gel without adding any further solution. The yellow dye migrates at about 40 bp leaving even short bands easily visible. Yellow SubTM has been successfully tested in PCR® applications using different Taq Polymerases and also other thermostable polymerases like Pfu, Tth and Pfx. Furthermore, Yellow Sub is even compatible with many restriction enzymes and is specially suited for restriction digest screening procedures. Yellow SubTM is successfully used in many laboratories and quoted in several recent studies.

Publications:
1. Pre-PCR Gel-Loading Buffer that Increases Specificity
Haack, K. Vizuete-Forster, M.
BioTechniques. 2000; 29:684-688.

2. Polymorphic Sex Testing
International Patent Application No. PCT/GB2002/003920; Filing Date: 27.08.2002

3. Tumor cells escape suicide gene therapy by genetic and epigenetic instability
Frank, O. et al.
Blood. 2004; Dec 1;104(12):3543-9

4. Epidemiological study of the prevalence of Babesia divergens in a
veterinary practice in the mid-east of France
Devosa, J. and Geysen, D.
Veterineray Parasitology. 2004; 125(3-4): 237-249.

5. Taeniasis and Cysticercosis in a Selected Group of Inhabitants from a Mountainous Province in Nother Vietnam
ITMA – MSTAH thesis, No 56, 2006.

6. Quantification of DNA methylation in electrofluidics chips (Bio-COBRA).
Brena RM, Auer H, Kornacker K, Plass C,
Nature Protocols. 2006;1(1):52-8.

7. Development and validation of a PCR-RFLP test to identify African Rhipicephalus (Boophilus) ticks
Laetitia Lempereur, Dirk Geysen and Maxime Madder
Acta Tropica, Volume 114, Issue 1, April 2010: 55-58

Product Features of Yellow SubTM

  • Increased yield in PCR® applications
  • Enhanced specificity in PCR® applications
  • Contains the loading buffer and dye - the reaction mix is applied directly to the agarose gel
  • Yellow dye migrates at 40 bp - even short PCR®-bands are easily visible
  • Compatible with all tested thermostable polymerases
  • Compatible with DNA-purification protocols (glass milk or Qiagen spin columns). PCR® fragments can be used in all regular applications like cloning or restriction digests
Figure 1. Yellow Sub does not inhibit Taq Polymerases. Taq Polymerases of 5 major manufacturers (M1 trough M5) were tested under standardized PCR®-conditions with (above) and without (below) Yellow Sub. In all cases, Yellow Sub did not inhibit the function of Taq Plymerases. In three cases (manufacturers 1, 3 and 5), specificity was significantly increased. All PCRs were performed in doubletts.
Lanes 1+2: Taq M1; lanes 3+4:Taq M2; lanes 5+6: Taq M3; lanes 7+8: Taq M4; lanes 9+10: Taq M5; lane 11: pUC Hpall DNA-Marker

Figure 2. Yellow Sub significantly increases specificity and yield. A temperature gradient ranging from 43,15 °C to 61,4 °C was applied under standardized PCR®-conditions with (above) and without (below) Yellow Sub. Under identical conditions, Yellow Sub enhanced the specificity and yield at all temperatures. Yellow Sub is very well suited for PCR® amplifications under which primer annealing temperatures are not well defined or DNA amplification leads to high amounts of unspecific products.
Lanes 1 through 9: Temperature gradient ranging from 61,4 °C to 43,15 °C; Lane 10: pUC Hpall

Product Application Protocols

  • Protocol 1: PCR®
        1. Prepare Reaction Mix
  • Thermostable DNA Polymerase 1 unit
  • 10 x Reaction Buffer 5µl
  • dNTP (10 mM) 1µl
  • Primer 1 0.1µM - 1µM
  • Primer 2 0.1µM - 1µM
  • Template DNA 10pg - 10µg
  • YellowSubTM 5µl
  • Add water to final volume of 50µl

2. Perform amplification reactions as usual
3. Agarose Gel Analysis
Load bewteen 5 - 50 µl of the reaction mix directly on the agarose gel. No additional loading buffer or loading dye is required! Alternatively, the PCR® products can be purified by column or glass milk purification or further enzymatic reactions (e.g. restriction enyzme digests) may be performed before Agarose gel analysis gel (i.e. DNA amplification followed by RFLP).
  • Protocol 2: Restriction digest
1. Prepare Reaction Mix
  • Restriction Enzyme 5-10 units
  • DNA 1µg
  • YellowSubTM 5µl
  • 10 x Reaction buffer 5µl
  • Add water to final volume of 50µl
2. Perform amplification reactions as usual

3. Agarose Gel Analysis
Load bewteen 5 - 50 µl of the reaction mix directly on the agarose gel. No additional loading buffer or loading dye is required.

Ordering Information

Quantity Order No. Price Euros* Pice USD  
5 x 1 ml Yellow Sub FL 115 79,- Euros 110,- USD  
10 x 1 ml Yellow Sub FL 120 130,- Euros 180,- USD  
50 x 1 ml Yellow Sub FL 130 520,- Euros 730,- USD  

* Prices plus 19% VAT for deliveries within the EU. Please provide VAT-ID if applicable.