Quantitative Protein Precipitation from Guanidine Hydrochloride-Containing Solutions by Sodium Deoxycholate/Trichloroacetic Acid
✍ Scribed by Ulrich Arnold; Renate Ulbrich-Hofmann
- Publisher
- Elsevier Science
- Year
- 1999
- Tongue
- English
- Weight
- 47 KB
- Volume
- 271
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
antisense primer, ER3A, at an annealing temperature of 61°C. The results are shown in Fig. 1C. Lanes 1-3 contain the PCR products generated with the targeted primer ER SX1/3 and the antisense primer ER3A using no DNA, pCRII-TOPO-2⌬, and pIC-ER-F as templates, respectively. Similarly, lanes 4 -6 contain the PCR products with the targeted primer, ER SX1/4, and the ER3A using no DNA, pCRII-TOPO-2-3⌬ and pIC-ER-F respectively as templates. As seen in Fig. 1C, the targeted primers designed for the exon 2⌬ and 2-3⌬ molecules amplified only the targeted splice junctions and generated the expected 414 bp and 296 bp products (lanes 2 and 5, respectively) but did not generate any products with the ER wild-type sequences (lanes 3 and 6, respectively). Thus the targeted primers which meet the requirement of at least three of four unique bases in the extreme 3Ј end exclusively amplify the alternate sequences. We believe that the principles developed in the current study with ER will have broad applicability to splice variants of a diverse range of genes.
Acknowledgments.