BC006 – Transformation of commercial bacteria with pGADT7-TRIM5a and -TRIM6 NEBuilder reactions

Submitted by Brittnee Crane

Commercial bacteria (C2987) was used to complete transformations for cloned samples: pGADT7-TRIM5α, pGADT7-TRIM6, pGADT7-TRIM69, pGADT7 empty (1), and pGADT7 empty (2). pGADT7-TRIM5α and pGADT7-TRIM6 successfully grew colonies, so LB broth was seeded to grow cultures intended for MiniPrep plasmid purification. Master plates were also made for the two samples. My partner, Hannah Sparks, completed the MiniPrep with the two cultures and ran the purified plasmid on a gel. The band that appeared on the gel for pGADT7-TRIM5α was the appropriate size, but the band for pGADT7-TRIM6 was not. She mixed a new PCR for TRIM6 using variant 2, instead of variant 1, in hopes that the appropriate amplicons would appear.

Leave a Reply

Fill in your details below or click an icon to log in:

WordPress.com Logo

You are commenting using your WordPress.com account. Log Out / Change )

Twitter picture

You are commenting using your Twitter account. Log Out / Change )

Facebook photo

You are commenting using your Facebook account. Log Out / Change )

Google+ photo

You are commenting using your Google+ account. Log Out / Change )

Connecting to %s