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<li>1 uL HindIII-HF</li> <li>1 uL SalI-HF</li>

<li>EcoRI-HF (NEB: 20000 U/mL)</li> <td class="tg-031e">EcoRI-HF</td>

...>a) Spytag-Streptavidin-ELP5-SnoopCatcher (to be cut with XhoI and HindIII-HF)</li> ...<li>a) HisTag-Snooptag-ELP5-Spycatcher (to be cut with XhoI and HindIII-HF)</li>

<th align="center">NotI-HF</th> <th align="center">EcoRI-HF</th>

<td class="tg-yw4l">EcoRI-HF<br></td> ...px;">12,3µl pYES2-CT (2ng) + 3µl Cutsmart + 12,7µl water + 1µl HindIII-HF + 1µl PmeI</div>

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<li>Performed RE digestion, nicked with EcoRI-HF (sample 1/2 stored in -20°C)</li> <li>EcoRI-HF Digested Sample 1 (D1)</li>

<td class="col0">EcoRI-HF</td><td class="col1">20U/µl</td><td class="col2">1</td> <td class="col0">EcoRI-HF</td><td class="col1">20U/µl</td><td class="col2">0.5</td>

<b>Digestion of insert and pSB1C3 with Pst1-HF and EcoRI-HF</b> Digestion of PCR products and psB1C3 with PstI-HF und EcoRI-HF in Cutsmart buffer

<b>Digestion of insert and pSB1C3 with Pst1-HF and EcoRI-HF</b> Digestion of PCR products and psB1C3 with PstI-HF und EcoRI-HF in Cutsmart buffer

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...NE buffer, 41µL of H2O at the end we added our enzymes. ( 2µL for ECORI HF and 2µL for PST1 ). ...:#cccccc;border:1pt solid #000000;padding:0.0694in;color:#000000;" | ECORI HF

<li>PCR product 4: BamHI (HF), NheI (HF) (NEB CutSmart) <li>PCR product 6: HindIII, NheI (HF) (NEB 2.1)

included preparing a mini-prep, mixing with enzyme BSr61-HF, enzyme KpuI-HF, buffer Cutsmart, and water. The remaining solution was then placed in incu <th>Kpn1-HF</th>

<td>AgeI-HF</td> <td>BamHI-HF</td>

0,5 ul EcoR1-HF (restriction enzyme)<br> 1ul EcoR1-HF (restriction enzyme)<br>

0.25 μl NcoI-HF<br/> 0.25 μl NcoI-HF <br/>

This is the Double Digest Protocol with EcoRI-HF and SpeI, using a common reaction and same incubation temperature for both Double Digest Recommendations for EcoRI-HF® + SpeI:

...et="_blank">PCR (Phusion polymerase) protocol</a> was followed. Instead of HF-buffer we used GC-buffer and DMSO.</p> <td>HF buffer</td>

<li>Buffer HF</li> <li>20 μl of 5X HF Buffer</li>