...cells by heatshock with the plasmid of interest. See our transformation page for a detailed heatshock transformation...

...introns. Avoid stretches of 4 or more nucleotide repeats, especially repeated Ts because polyT is a termination...an ampicillin analog). Due to the long terminal repeats found in lentiviral plasmids, Addgene currently...or NEB Stable cells. This will ensure that the repeats will be maintained and often results in a greater...

...Disinfectant Thermometer Water bath weights and floats Reagents None needed Procedure Ensure that the ...water bath when they are placed inside. Water bath floats can be used to secure tubes in place and water ...

...prior to handling the materials. Equipment Weighing boats or weighing paper Balance Scoopula or spatula Container...

...Link opens in a new window) OpenWetWare: Janet Matsen’s guide to Gibson Assembly (Link opens in a new ...

...swirling the flask occasionally as the solution heats up.). Caution HOT! Be careful stirring, eruptive...

...Denaturation for 2 minutes at 94°C: This initiation step heats the double stranded DNA template strand to the point...