IG Participation Form

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Requested material for accreditation

Shipment

If shipment by transportation paid by the participant, please provide name of company

Source of cells

Anticoagulant

Work-up of cells

Nucleic Acid

DNA Extraction Method

RNA Extraction / cDNA Synthesis Methods

General Strategy

5´ Primer Used

3´ Primer Used

Reference for Primer Sets Used

Quantity of DNA / cDNA Used

Taq Polymerase Used (Including Company Name)

Analysis of PCR Products Prior to Sequencing

If yes, by

Purification of PCR Product Prior to Sequencing

Cloning of PCR Product Prior to Sequencing

If yes, please provide the situations and estimated frequency for when cloning is applied

Cloning Kit Used (including Company Name)

Prior to sequencing, do you check the clones for the correct insert?

Number of clones selected for Sequencing

General Strategy

Primers Used for Sequencing

Strands Sequenced

Bioinformatics Tools for Sequence Analysis - Please specify tools/programs used for: Visualisation of Chromatograms

Please specify tools/programs used for: Alignment of Both Strands (Forward and Reverse)

Please specify tools/programs used for: Multiple Sequence Alignment e.g. after Cloning

Please comment on any additional bioinformatic databases utilised

Bioinformatics Tools for Sequence Interpretation - Please specify the Tools/Programs utilised for each of the following: Identification of IGHV, IGHD, IGHJ Gene and Allele

Calculation of the Percentage Identity to the Germline

Do you exclude from the calculation the sequence corresponding to the 5´ primer?

Do you include in the calculation mutations within the HCDR3 of the IG gene?

Interpretation of Difficult Cases: Briefly outline how each of the following situations are handled/interpreted: Single Unproductive Rearrangements

Double Productive Rearrangements

Briefly outline the information you currently provide in a Laboratory Report for IGHV Gene Mutation Status, e.g. tissue type, gDNA/cDNA, PCR primers, gene usage, % identity, functionality, conclusion, etc.

Briefly outline how you would report each of the following cases: 1. Mutated ´Borderline´ Rearrangement

2. Single Unproductive IGHV Rearrangement

3. Double Productive IGHV Rearrangements

4. Double Productive Rearrangements with Discordant Mutational Status

5. IGHV3-21 Rearrangement

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