Server at Davidson College

Prime Time Primer Designer
A Golden Gate Assembly Tool

Instructions
Sources

*Please enter the plasmid sequence (plain text):

*Please enter the sequence you wish to insert:

*Restriction Enzyme:	BsaI
*Primer length:
*Starting location of cut:
*Ending location of cut:

*Choose two sticky ends (one letter and one number):

	A				1
	B				2

* = required

 

1. Plasmid Sequence

   Insert the text of the plasmid sequence. Only plain-text is supported. New-line characters will be automatically deleted.

2. Insert Sequence

   Insert the sequence you wish to insert. Only plain-text is supported. New-line characters will be automatically deleted.

3. Restriction Enzyme

   Select the restriction enzyme you will use from the drop down menu. Currently, BsaI is the only supported restriction enzyme. The restriction enzyme is used to generate the specific tag (i.e. GGTCTC).

4. Melting Temperature

   Insert the desired melting temperature. The default value is 50.0°C. If two primers are greater than 5.0°C apart, the one with the lower melting temperature will be elongated until they are within 5°C.

5. Primer Length

   Insert the desired primer length. Default value is 20. If melting temperatures differ, the primer length may be altered (elongated) to make the melting temperatures more similar.

6. Starting Index

   Insert location of the start of the cut. This index should be the last base pair that you want included. Indices start at "1"

7. Ending Index

   Insert location of the end of the cut. This index should be the first base pair that you want included. Indices start at "1"

8. Sticky Ends

   Choose the two sticky ends you wish to use. Note that only one letter and one number may be selected.

9. Submit

   That's it! Submit and collect your results.