Home FAQs What is the difference between the original and version 2 (v2) Phage Display kits?

FAQ: What is the difference between the original and version 2 (v2) Phage Display kits?

The exact differences between the original version and the v2 kits are outlined in the table below.  The Ph.D.-7, Ph.D.-12 and Ph.D.-C7C libraries have not changed, and their product numbers remain the same.  The new v2 kits include a streamlined and modernized solution-phase panning control experiment that requires fewer experimental steps, whereas the original kits recommend a surface panning experiment. Additionally, the -96 gIII Sequencing Primer is now supplied at a 10-fold higher concentration (10 pmol/ml) to better align with commercial sequencing facility submission guidelines. The -28 gIII Sequencing Primer has been removed because the annealing location is too close to the Ph.D. random region to be useful in modern sequencing technologies.

Original Kit Components
Name Product # Amount Concentration
Ph.D. Phage Display Peptide Library E8102 or E8111 or E8121 1 x 0.1 ml 1 x 1013 pfu/ml
E. coli K12 ER2738 E4104 1 x 0.2 ml
-96 gIII Sequencing Primer S1259 100 pmol 1 pmol/ml
-28 gIII Sequencing Primer S1258 100 pmol 1 pmol/ml
Streptavidin N7023 1 x 1.5 mg
Biotin N7024 1 x 0.1 ml 10 mM

New v2 Kit Components
Name Product # Amount Concentration
Ph.D. Phage Display Peptide Library E8102 or E8111or E8121 1 x 0.1 ml 1 x 1013 pfu/ml
E. coliK12 ER2738 E4104 1 x 0.2 ml
-96 gIII Sequencing Primer S1259 500 pmol 10 pmol/ml
DYKDDDDK Mouse mAb E8004 1 x 0.015 ml
Protein G Magnetic Beads S1430 1 x 0.15 ml

Original Kit Method

New v2 Kit Method

 

Surface Panning

Solution Phase Panning

Capture

Polystyrene surface (well or petri dish)

Protein G Magnetic Beads (#S1430)

Prepare Capture Surface

Overnight coating of target (Streptavidin, #N7023), 2 hr blocking

None

Binding (Selection Step)

Apply Ph.D. library to coated surface (10-60 min, RT)

Mix target (DYKDDDDK Mouse mAb, #E8004) + Ph.D. library (20 min RT). Capture phage-target complexes with Protein G Magnetic Beads (#S1430)

Wash

10 x 1ml TBST, pipet or dump

10 x 1ml TBST, pellet with magnet

Elution

1 mL of 0.1 mM Biotin (#N7024) (30 min RT)

1 mL of pH 2 Glycine Buffer (10-20 min RT), neutralize to pH 9 with TrisHCl

Enrich in E. coli culture (4-5 hr)

Repeat selection 1-2 more times

Sequencing and/or binding studies (-96 gIII Sequencing Primer, #S1259)