In silico cloning - Bits.vib.be

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This will add an annotation to this region, Forward primer region, (unselect Show in the Restriction sites settings in the Side Panel to get a more clear view of the annotations) and 5 rows of red and green dots (you have to zoom in to see the dots: use the 100% button) representing the primer suggestions. There is a line for each primer length - 18bp through to 22 bp, hence five lines. 1.2 Examining the primer suggestions Each line consists of a number of dots, each representing the starting point of a possible primer, e.g. the first dot on the first line represents a primer starting at the dot's position and with a length of 18 nucleotides. The primer will anneal to the selected region in the DNA. Position the mouse cursor over a dot. A box will appear, providing data about this primer.
Clicking the dot will select the region where that primer would anneal. Note that some of the dots are colored red. This indicates that the primer represented by this dot does not meet the requirements set in the Primer parameters that you can see in the Side panel on the right: The default maximum melting temperature is 58. This is the reason why the primer in the figure below with a melting temperature of 58.55 does not meet the requirements and is colored red. There is an asterisk (*) before the melting temperature. This indicates that this primer does not meet the requirements regarding melting temperature. In this way, you can easily see why a specific primer fails to meet the requirements.
Page 1: Tutorial 1: Primer design In this t
Page 5 and 6: By adjusting the Primer parameters
Page 7 and 8: In the Side Panel of the view conta
Page 9 and 10: Tutorial 2: In silico cloning workf
Page 11 and 12: You can do all kinds of restriction
Page 13 and 14: Click Next In this step you specify
Page 15 and 16: Click OK and the sequence will be i
Page 17 and 18: fine the minimum number of position
Page 19 and 20: Use the Browse button to select pcD
Page 21 and 22: Next step is to cut the fragment. A
Page 23 and 24: Save the construct shown in the cir
Page 25 and 26: Switch to the fragment sequence. Cl
Page 27 and 28: Note that the Workbench is validati
Page 29 and 30: 4.1 Add attB sites The first step i
Page 31 and 32: This will allow you to choose which
Page 33 and 34: You also get a list of primers. Aga
Page 35 and 36: Next you have to specify the entry
Page 37 and 38: When you have selected your entry c

In silico cloning - Bits.vib.be

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