Cloning Request Form for Mutagenesis
(PROTEIN EXPRESSION LABORATORY, Enquiries on ext. 7202/7092)
NAME:
Under/graduate/Postdoc/PI
CHARGE CODE:
DEPARTMENT:
PI’s NAME:
LAB No:
DATE:
Email Address:
TEL (Ext.):
Primary PCR
Clone Name
PCR Name
Primer 1
Primer 2
Template Name
Template
PCR
Template Secondary
PEL No.
Resistance
Length
PCR
(PEL Use only)
Cloning Request Form for Mutagenesis
(PROTEIN EXPRESSION LABORATORY, Enquiries on ext. 7202/7092)
NAME:
Under/graduate/Postdoc/PI
CHARGE CODE:
DEPARTMENT:
PI’s NAME:
LAB No:
DATE:
Email Address:
TEL (Ext.):
Secondary PCR and Cloning
No
Clone Name
Primer 1
Primer 2
Template
PCR Template
PCR Insert
pLEICS
Known Toxicity
Verification
Name
Resistance
Length (bp)
Vector Name
to E. coli strains
Method
Protex No.
Verification Method:
A: Colony PCR (basic rate, no free repeats) £35.00 per mutagenesis with PCR verification
B: Sequencing (no-win-no fee-rates, turnaround time: 3 weeks)
Rates: £120.00/clone inserts ≤1000bp; £150.00/clone, inserts ≤2000bp; £250.00/clone, inserts ≤3000bp (insert sequence files needed)
Materials requirement: 1. PCR Template (50-200ng/μl, 8μl/reaction) 2. Primers (1μg/μl or 10-50μM, 8μl/reaction)
3. PCR products (gel purified 8μl)
Please can you confirm that this work has been (or is in the process of being) approved by the GM subcommittee? Yes , GM Reg.No.
If no, it is your responsibility to contact Dr. Christopher Bruce x2020 as soon as possible
Is there any particular hazard associated with this insert? e.g. is it oncogenic or toxic.
Yes
No
No
If clones are for the collaborators outside the University of Leicester, please give details.
SUCCESS RATES:
Overall our success rate is >90%. However, please note that success rate can vary depending upon a number of factors:
1. Which company supplies the primers (for advice contact xy13)
2. Insert length (inserts ≥3000bp or ≤100bp are troublesome)
3. Which particular pLEICS vector is being cloned into (high copy number work best)
4. The gene being cloned (some genes are toxic)
Please let us know if sequenced clones are not correct. Otherwise, we shall assume that they are correct. This feedback is key to help us improve the service.