Extension Time In Pcr In Georgia
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You do NOT need to request an extension to file your Georgia return if you receive a Federal extension. The due date for filing your Georgia return will be automatically extended with an approved Federal extension.
Request an additional six months to file your Georgia income tax return. There are two ways to request an extension to file your Georgia income tax return. You can either request an extension on your federal return or fill out Georgia Form IT-303 if you only need a state extension and don't need a federal extension.
You must be 65 years old or older. You must be living in the home to which the exemption applies on January 1 of the year for which the exemption applies. Your net income, or the combined net income of you and your spouse must not be greater than $10,000 for the preceding year.
Georgia Law provides 45 days for a property owner to file a written appeal. Realizing that a county must complete work on the annual digest by July 1, and submit it to the Department of Revenue by August 1, the law does not provide for extensions to this deadline.
Excessive extension time can allow nonspecific amplification. Generally, use an extension time of 1 min/kb. Excessive annealing time may increase spurious priming. Use an annealing time of 30 sec.
Extension Time Extensions are normally performed at 68°C. As a general rule, use extension times of one minute per 1000 base pairs (e.g. 3 minutes for a 3 kb product) For products less than 1 kb, use 45-60 seconds. Products greater than 3 kb, or reactions using more than 30 cycles, may require longer extensions.
Increase the annealing temperature to improve specificity. The optimal annealing temperature is usually no less than 3–5°C below the lowest primer Tm. Optimize the annealing temperature stepwise in 1–2°C increments, using a gradient cycler when available. Consider touchdown PCR to enhance specificity.
Extension is achieved by using the loosened nucleotides of each base to grow the complementary DNA strand. The end result is two double-stranded products of DNA. The temperature that is used during the extension phase is dependent on the DNA polymerase that is used.
If the extension time is too short, there will be insufficient time for complete replication of the target. Generally, use an extension time of 1 min/kb. If the annealing time is too short, primers do not have enough time to bind to the template. Use an annealing time of at least 30 sec.
The extension time of PCR depends upon the synthesis rate of DNA polymerase and the length of target DNA. The typical extension time for Taq DNA Polymerase is 1 min/kb, whereas that of Pfu DNA polymerase is 2 min/kb.
