How to Design Primers for PCR

Oligonucleotide primers are necessary when running a PCR reaction. One needs to design primers that are complementary to the template region of DNA. They are synthesized chemically by joining nucleotides together. One must selectively block and unblock repeatedly the reactive groups on a nucleotide when adding a nucleotide one at a time. The main property of primers is that they must correspond to sequences on the template molecule (must be complementary to template strand). However, primers do not need to correspond to the template strand completely; it is essential, however, that the 3’ end of the primer corresponds completely to the template DNA strand so elongation can proceed. Usually a guanine or cytosine is used at the 3’ end, and the 5’ end of the primer usually has stretches of several nucleotides. Also, both of the 3’ ends of the hybridized primers must point toward one another.
___________________________________________________________________________
Chapters:
0:00 Introduction to Primer Design
0:57 Primer Length
1:26 Annealing & Melting Temperatures
2:27 Makeup of Nucleotides in the Primer
3:20 Secondary Structure
4:04 You Just Designed a Primer!

___________________________________________________________________________
Credits
Featuring: Jennifer Tsang
Shot by: Jennifer Tsang
Written by: Jennifer Tsang & Quintin Marcelino
Directed, Animated, & Edited by: Quintin Marcelino
Music by: Lauren Duski, Vibe Mountain, & RKCV

[Nini]

I like the way you explained everything with total calm and understanding, keep going and thank you❤

[samar]

Thank you sm. They just explained this to us in class and I didn't understand anything until I watched your video.

[yvius]

Thank you so much for this video! It's very helpful and explains the primers very well. And it'll surely help for my biotechnology exam next week^^

[deniz]

Thank you very much! It is well-explained!! I never figured out the basic PCR design until I looked at you! You're awesome!! :)

[bakhtiyorsheraliev]

Wonderful explanation! I hope you keep update.

[ramkomusique]

Thanks! This is a super neat introductory video for primer design :)

[a.s]

What are the online tools to warn us of potential hairpins, self-dimers, and cross-dimers?

[shreyaagarwal]

Complete information in the most simplified manner

[caveman]

Thanks for this video! i gotta design primers for my project and this has helped me so much :))

[bradhilton]

Thanks Jennifer, That was both interesting and informative .

[adamquazar]

Thank you, Miss Ma'am! I love this video of yours, so comprehensive!

[anns]

I did that lil dance at the end with you 💃✨

[susandoming]

What if you don’t know the DNA sequence? How do you choose primer?

[HaloBro]

This has been really helpful, thank you so much!

[Sanariyadlier]

the content was great and so you were so sweet love it

[solmazshirjang]

It would be great if you could make a video about CRISPR KI... There is very little information available about this technique.

[ddkisa]

Omg, this is gold. Thank you so much:)

[omdah]

My advice for y’all guys always use softwares when designing your primer will save you more time and much efficient .

[carlydelsack]

helps a bunch!!! very concise and clear!

[monimakarmacharya]

Thank you so much it really helped a lot. I have my genetic engineering exams in few days. 😇😇😇✊🏻✊🏻