In silico cloning - Bits.vib.be

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Select the ATP8a1 mRNA sequence | Click Next Use the Browse button to select the two primer sequences | Click OK Leave the other parameters at their default values. The Match criteria determine how well a primer should match the sequence. Exact match specifies that all positions of the primer must base pair with the sequence. Note that we cannot choose this option because we have added non-matching restriction sites to the primers. We have to de-
fine the minimum number of positions and the number of consecutive positions in the 3’ end of the primer that must base pair to the sequence in order to speak of a match. Note that the number of mismatches is reported in the output, so you can filter on this afterwards. Below the Match criteria, you can adjust Concentrations concerning the reaction mixture. This is used when reporting melting temperatures for the primers. Click Next | Adjust the output options Click Finish You now see the fragment table displaying the PCR product. In the Side Panel you can choose to show information about melting temperature for the primers. Right-click the fragment and select Open Fragment.
Page 1 and 2: Tutorial 1: Primer design In this t
Page 3 and 4: Clicking the dot will select the re
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: Click OK and the sequence will be i
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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