Primer Synthesis 10 nM 15-34bp
Price may change significantly. Please contact for details and discount.
| Technical | |
| Technical Details | Details The 2-step PCR is the first step in the protocol of CFPS700 kit to generate the DNA template to be used in the following transcription and translation. The 1st PCR aims to connect the target ORF sequence with the 2nd forward/reverse primers (please see the diagram below). Therefore, the 1st PCR forward/reverse primers need to be target gene specific that overlap with the N-terminal and C-terminal sequences of the ORF of the target gene. There are two sets of forward and reverse primer sets in the 2nd PCR. The two reverse 2nd PCR primers and one forward 2nd PCR primers are universal primers. The other forward 2nd PCR primer contains a tag sequence, such as His-, FLAG-, or MYC-tag, therefore can differ from one another depending on your tag choice. In the CFPS700 protocol, overlapping length between 1st gene specific primers and ORF is set at 18 bp as the default. But the overlapping length can be extended to 30 bps, giving multiple options for primer. For some options, depending on the GC content of N- or C-terminal sequences of ORF, Tm value difference between one 1st gene specific forward primer candidate and 1st gene specific reverse primer candidate can be higher than optimal. When this happens, the selection of this primer set would result in less efficient or even failed 1st PCR, subsequently 2nd PCR, and ultimately failed protein translation. Therefore, carefully selecting the optimal 1st PCR primer set with the minimum Tm value difference is very important to the success of both the 1st and 2nd PCRs and ultimately protein translation using CFPS700 kit Reference: https://www.sigmaaldrich.com/technical-documents/articles/biology/instruction-for-the-primer-design-tool-for-the-1st-pcr.html?gclid=CjwKCAjw7-P1BRA2EiwAXoPWA_PO4xZm7g-5WVxiLPHaJ-99pJ1M-2C6OY8X2fo8Lk4gHBrZGDjGIhoCLOMQAvD_BwE |
| Protocol | |
| Protocol 3 | Purpose of developing the primer design tool for the 1st PCR The Primer Design Tool for the 1st PCR is available to download at the bottom of the page. The 2-step PCR is the first step in the protocol of CFPS700 kit to generate the DNA template to be used in the following transcription and translation. The 1st PCR aims to connect the target ORF sequence with the 2nd forward/reverse primers (please see the diagram below). Therefore, the 1st PCR forward/reverse primers need to be target gene specific that overlap with the N-terminal and C-terminal sequences of the ORF of the target gene. There are two sets of forward and reverse primer sets in the 2nd PCR. The two reverse 2nd PCR primers and one forward 2nd PCR primers are universal primers. The other forward 2nd PCR primer contains a tag sequence, such as His-, FLAG-, or MYC-tag, therefore can differ from one another depending on your tag choice. |
Primer Synthesis
- Brand: Bioport
- Product Code: 2000418
- Reward Points: 600
- Availability: In Stock
-
₹18
- Ex Tax: ₹18