blob: b5f58d5c0ec2e1dc2eb72e721de105eca5335356 (
plain)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
|
# from cathal
This was from Cathal Garvey a while back:
(02:43:38 PM) Cathal Garvey: Here, this enzyme: Terminal Transferase and easily removed blocking groups on NTP washes = Easy homebrew DNA synthesis reaction? http://en.wikipedia.org/wiki/Terminal_deoxynucleotidyl_transferase
(02:44:13 PM) Cathal Garvey: In other words, not so dissimilar to the existing method, but without requiring the chemistry of actually joining nucleotides with such care. In fact, you could probably do something like this:
(02:44:21 PM) Cathal Garvey: Oligo annealed to solid substrate
(02:44:29 PM) Cathal Garvey: Ends not phosphorylated at beginning of each step.
(02:45:09 PM) Cathal Garvey: Add a kinase to prime DNA, add NTP + TDT
(02:45:32 PM) Cathal Garvey: Interesting stuff!
(02:45:44 PM) Cathal Garvey: I love all the little methods devised by nature for this function
(02:45:49 PM) Cathal Garvey: By the way, my error there:
(02:46:06 PM) Cathal Garvey: the method I suggest is based on the assumption that the enzyme might bind the oligo end and await an NTP,
(02:46:16 PM) Cathal Garvey: and that you can use NDPs rather than NTPs
(02:46:35 PM) Cathal Garvey: so that it won't polymerise further than a single nucleotide without a step involving kinase
(02:47:06 PM) Cathal Garvey: So you can use two enzymes and NDPs in washes, if that prior assumption regarding the enzyme holds.
(02:47:15 PM) Cathal Garvey: Which is, of course, a huge achilles heel.
## more recent
This is from Cathal more recently: "I have a concept that's only use enzymes, DNA and triphosphates. Hoping to try it someday soon for a POC."
# related igem projects
* [oligonucleotide synthesis using Terminal Deoxynucleotidyl Transferase (TdT) as an alternative to phosphoramidite-based DNA synthesis](http://2014.igem.org/Team:Cooper_Union/TdT_project)
* [DNA synthesis: characterization of Terminal Deoxynucleotidyl Transferase as an alternative oligonucleotide synthesis method](http://2014.igem.org/Team:Rutgers)
|
