Primer Design: Important Considerations and Tips for Good Primer Design

Running a PCR experiment and need to design primers? We all have been there. Primer design is essential for successful PCR, but where do you start? In this video, we’ll go through important considerations and tips for good primer design!
Here are some helpful time stamps for our Primer Design video:
0:41 Primer Length
1:06 Considerations for Melting Temperature (Tm) and Annealing Temperature (Ta)
2:40 GC Content
3:16 Avoid Repeats in Your Primer Sequence
3:39 Avoid Complementary Sequences Within the Primer
4:06 Avoid Secondary Structures in Target Sequences
4:31 Summary
5:07 Avoid List
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[@abmgood]

Here are some helpful time stamps for our Primer Design video:
0:41 Primer Length
1:06 Considerations for Melting Temperature (Tm) and Annealing Temperature (Ta)
2:40 GC Content
3:16 Avoid Repeats in Your Primer Sequence
3:39 Avoid Complementary Sequences Within the Primer
4:06 Avoid Secondary Structures in Target Sequences
4:31 Summary
5:07 Avoid List

[@bio366geethasankar7]

🙏🏻

[@bytesizebiotech]

What is the reason for no more than 3x GCs in the last 5 bps? I've never had an issue with doing so. I always read that it's "At least 3, and up to 5 base pairs"

[@abmgood]

The reason to prevent having more than 3x GCs is to lower the chance of self clamping on the 3' end. Thank you for sharing your success with our viewers!

[@polinaskrobova5636]

Hi! What secondary annealing means?

[@faanutk7861]

hairpin and selfdimer are secoundry annealing

[@tayyabanazim100]

❤

[@MohammadMahmoud-ji1jc]

I have created arms PCR primer; might I perhaps email it for revision?

[@abmgood]

Yes of course! Please email your inquiry to [e-mail], and our tech team will look into it! :D

[@MohammadMahmoud-ji1jc]

@@abmgood thank you

[@nurulainafandi4267]

What is meant by Tm of product?...what is meant by word product?????

[@alfomeg588]

melting temperature